{"paper_id":"25d3a93f-a897-4f17-b771-66c004e9f9b4","body_text":"Characterization of immortalized ovarian epithelial cells with BRCA1/2 mutation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Characterization of immortalized ovarian epithelial cells with BRCA1/2 mutation Hiroaki Komatsu, Masayo okawa, Yasuhiro Kazuki, kanako Kazuki, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4012283/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 14 Apr, 2024 Read the published version in Human Cell → Version 1 posted 4 You are reading this latest preprint version Abstract We aimed to elucidate the mechanism underlying carcinogenesis by comparing normal and BRCA1/2- mutated ovarian epithelial cells established via Sendai virus-based immortalization. Ovarian epithelial cells (normal epithelium: Ovn; with germline BRCA1 mutation: OvBRCA1; with germline BRCA2 mutation: OvBRCA2) were infected with Sendai virus vectors carrying three immortalization genes ( Bmi-1 , hTERT , and SV40T ). The immunoreactivity of anti-epithelial cellular adhesion molecule (EpCAM) antibodies in each cell line and cells after 25 passages was confirmed using flow cytometry. Chromosomes were identified and karyotyped to detect numerical and structural abnormalities. Total RNA extracted from the cells was subjected to human transcriptome sequencing. Highly expressed genes in each cell line were confirmed using real-time polymerase chain reaction. Immortalization techniques allowed 25 or more passages of Ovn, OvBRCA1, and OvBRCA2 cells. No anti-EpCAM antibody reactions were observed in primary cultures or after long-term passages of each cell line. Structural abnormalities in the chromosomes were observed in each cell line; however, the abnormal chromosomes were successfully separated from the normal structures via cloning. Only normal cells from each cell line were cloned. MMP1 , CCL2 , and PAPPA were more predominantly expressed in OvBRCA1 and OvBRCA2 cells than in Ovn cells. Immortalized ovarian cells derived from patients with germline BRCA1 or BRCA2 mutations showed substantially higher MMP1 expression than normal ovarian cells. However, the findings need to be validated in the future. ovarian epithelial cancer carcinogenesis Sendai virus BRCA1/2 Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction The pathogenesis of ovarian cancer is becoming clearer in terms of the histological and clinicopathological background. A recent study has shown that the origin of ovarian cancer, especially high-grade serous carcinoma, is serous tubal intraepithelial carcinoma [ 1 ]. However, analysis at the molecular level remains insufficient [ 2 ], which may be attributed to the lack of sufficient cell lines derived from normal ovarian epithelium, benign lesions, and borderline malignant lesions to allow for a detailed analysis of the mechanisms involved in malignant transformation. BRCA1 and BRCA2 are crucial genes involved in ovarian cancer development, and germline mutations in these genes have been linked to a high frequency of breast and ovarian cancers [ 4 , 5 ]. However, there are no reports on predictors or appropriate diagnostic markers for cancer development in patients with BRCA1 and BRCA2 mutations, making regular examination and measurement of tumor markers crucial. Although BRC A mutations in mammary epithelial cells have been related to genetic instability, studies on BRCA in immortalized cells are intriguing [ 5 ]. Establishing normal ovarian epithelial cells and cells with BRCA1 /2 mutations will considerably expand the scope of research, including the analysis of genes involved in carcinogenesis, factors that inhibit carcinogenesis, and mechanisms underlying carcinogenesis. However, ovarian epithelial cells cannot be passaged over a long period and must be collected and cultured each time as research material. This process is difficult because ovaries are intraperitoneal organs. Sendai virus (SeV) vectors have a genome of single-stranded minus-strand RNA that can be introduced into non-dividing and dividing cells and have a high ability to express foreign genes [ 6 ]. As the vector genome remains in the cytoplasm, it does not affect host chromosomes and does not induce structural changes or mutations; therefore, it has been used as a tool to identify the causes of rare diseases and devise suitable treatments [ 7 – 10 ]. In the present study, we attempted to establish immortalized normal ovarian epithelial cells and ovarian epithelial cells from patients with germline BRCA1 or BRCA2 mutations using SeV vectors and to confirm whether the properties of each cell line could be analyzed. Materials and Methods Patient background Patient A was 49 years old and underwent total hysterectomy and bilateral ovarian and fallopian tube resection for atypical endometrial hyperplasia; the epithelium of the ovarian surface layer was extracted via a fine surgical cut using a scalpel. Patient B, aged 57 years, developed breast cancer, and genetic testing revealed a BRCA1 mutation (C.405_406 del (P.Arg 136 Thrfs*5)) and was diagnosed with hereditary breast/ovarian cancer (HBOC). Patient C developed breast cancer at the age of 53 years, and genetic testing revealed a BRCA2 mutation (C.1813 del (P.lle 605 Thrfs*9)) and was diagnosed with HBOC. In Patients B and C, the ovarian epithelium was collected via abrasion from surgical specimens using risk-reducing salpingo-oophorectomy (RRSO). In none of these cases of ovarian or fallopian tube carcinoma, the serous tubal intraepithelial carcinoma (STIC) was grossly or histopathologically observed in hematoxylin and eosin (HE)-stained specimens with paraffin block embedding. Establishment of primary cell lines The surface of the excised ovarian tissue was cut into small pieces with a scalpel, and 10 mL of 1% collagenase + Dulbecco’s modified Eagle’s medium (DMEM)/Ham’s F-12 with l -glutamine and sodium pyruvate (Product Number: 045-30665) was added to a 15 mL tube. The tube was incubated horizontally in a 37°C incubator (100 rpm) for approximately 24 h. Subsequently, the mixture was filtered twice into a 50 mL tube using a 70 µm strainer (Product Number: 352350). The mixture was then centrifuged at 1,300 rpm for 8 min, and the supernatant was discarded. The cells were washed with 2 mL of DMEM/Ham’s F12, and then 8 mL of DMEM/F12 was added; the mixture was transferred to a 15 mL tube and filtered into a 50 mL tube using a 40 µm (Product Number: 352340) strainer. The cells were seeded in a 60 mm dish with DMEM/Ham’s F12 + 20% fetal bovine serum (FBS) + 1.0 penicillin–streptomycin and incubated in a CO 2 incubator at 37°C. The next day, blood cells were removed by washing with phosphate-buffered saline (PBS), and the cells were seeded at a density of 2.0 × 10 5 cells/well in a 24-well plate. On the following day, the cells were infected with an SeV vector carrying three immortalization genes ( hTERT , Bmi1 , and SV40T ). The SeV vectors were used for gene loading and vector production by ID Pharma, Inc. Finally, the infected cells were subjected to long-term passaging. SeV vector infection The cells were seeded (2 × 10 5 cells) in 24-well plates. The next day, the cells were infected with SeV-hTERT (with simultaneous enhanced green fluorescent protein (EGFP) loading), SeV-Bmi1 (with simultaneous orange fluorescent protein (OFP) loading), and SeV-SV40T in a safety cabinet (the amount of virus used for infection is not disclosed owing to a pending patent). Flow cytometry The cell count was 2.5 × 10 5 cells/100 mL PBS + 1% FBS/well. Immortalized cells derived from human tissues (Ovn Passage + 20, Ov BRCA1 Passage + 5, Ov BRCA1 Passage + 20, Ov BRCA2 Passage + 20) were used along with positive control cells (colon cancer cell line HCT116, ovarian clear cell carcinoma cell line TU-OC-1 [ 11 ], and ovarian serous adenocarcinoma cell line TU-OS-6). Flow cytometry was performed with accutase treatment for 1–3 min when the confluency of all cell lines reached approximately 70% (Online Resource 1). The survival rate was approximately 90% for all cell lines. The primary antibodies (1 mg/100 mL/well) used were anti-EpCAM antibody (hZAK3A101; original stock: 1.105 mg/mL; 20210505) and negative control antibody (hIgG; original stock: 1 mg/mL; Invitrogen; 12000C; S1257958). The cells were incubated at 4°C for 1 h and then washed twice with PBS + 1% FBS (200 mL/well). The secondary antibody anti-hIgG antibody Alexa 488 (original stock: 1.5 mg/mL; Jackson ImmunoResearch, 109-545-190) was added (0.075 mg/30 mL/well) to the cells, and the mixture was incubated at 4°C for 1 h and then washed twice with PBS + 1% FBS (200 mL/well). Cloning of ovarian cells The established cell lines were seeded at low concentrations in a 100 mm dish. The cells were observed for more than 40 days, and the area where colonies formed from single cells was observed using an all-in-one microscope to confirm that both green fluorescent protein (GFP) and OFP were detected. The cells were selected under a microscope and seeded in 96-well plates. This step was repeated multiple times to pass on more than 50 cloned cells, scaling up from 96-well plates to 24-well plates, 12-well plates, 6-well plates, 60 mm dishes, and 100 mm dishes, in that order. Cells that could be passed on were designated as cloning cells. The cloned cells were defined as those that could be passaged. Chromosome analysis Each cell line was seeded in a 60 mm dish, treated synchronously with MAS (Genial Genetic Solutions), and then fixed using the Carnoy’s fixation method. The fixed cells were spread on slides and stained with quinacrine mustard (QH; Sigma–Aldrich, Hoechst 33258; Sigma–Aldrich) for multicolor fluorescence in situ hybridization (mFISH) (24XCyte: MetaSystems Inc., Altlußheim, Germany), according to the manufacturer’s instructions (MetaSystems). Metaphase images were captured with an AxioImagerZ2 fluorescence microscope (Carl Zeiss GmbH, Jena, Germany) and analyzed using the Ikaros software program (MetaSystems) and ISIS software program. Chromosome karyotyping of metaphase was performed to determine the chromosome number and karyotypic structural aberrations. Human transcriptome sequencing analysis Total RNA was extracted using the RNeasy Mini Kit (Qiagen; cat. No. 74104, 74106), and quality control was performed using the Agilent 2200 TapeStation to confirm that there was at least 2.0 µg of RNA. Analysis was performed using NovaSeq 6000 (Illumina). Libraries were prepared using the TruSeq stranded mRNA Library Prep Kit with a library length of 332–346 bp. The read length was 101 bp and was analyzed using the multiplex method. Cluster analysis was performed using the k-means method by classifying clusters into five categories according to the slope of the expression variation ratio. Real-time polymerase chain reaction Total RNA was extracted from each cell line using the RNeasy Mini Kit (Qiagen; Cat. Nos. 74104 and 74106). DNase treatment was performed using deoxyribonuclease (RT Grade, Cat. No. 313–03161) and the RNaseOUT™ Recombinant Ribonuclease Inhibitor (Cat. No. 10777019). A high-capacity cDNA Reverse Transcription Kit (Applied Biosystems; Cat. No. 4368814) was used for cDNA synthesis. Real-time polymerase chain reaction (PCR) was performed using the StepOnePlus Real-Time PCR system (Applied Biosystems) and TaqMan Fast Advanced Master Mix (Applied Biosystems) according to the manufacturer’s protocol. Transcript levels were normalized to those of beta-actin (ACTB). The PCR conditions were as follows: 50°C for 2 min, 95°C for 20 s, followed by 40 cycles at 95°C for 1 s and 60°C for 20 s. Each reaction was performed in triplicate. The PCR primer sequences are shown in Online Resource 2. PCR primers that showed more than 2-fold gene expression in Ov BRCA1 and Ov BRCA2 compared with that in Ovn upon human transcriptome sequencing analysis were selected. Ethics statement All patients provided written informed consent following the institutional guidelines. All methods were carried out in accordance with relevant guidelines and regulations. The study was approved by the Ethics Committee of Tottori University (IRB number: 20A179). Results Morphological and growth characteristics Ovn, Ov BRCA1, and Ov BRCA2 exhibited spindle-shaped or similar round cell morphology. All cell lines showed GFP and OFP fluorescence (Fig. 1 ). The growth curves for each cell type are shown in Fig. 2 . Each SeV-infected cell line showed growth, and SeV-uninfected Ovn, Ov BRCA1, and Ov BRCA2 cells survived for up to 10, 20, and 35 days, respectively; however, no further growth was observed. Temperature dependence For Ov BRCA1, the temperature was increased from 35°C to 37°C in the CO 2 incubator. The intensity of GFP and OFP staining decreased markedly on day 10 after the temperature change (Online Resource 3), and the cells showed no growth after 20 days of temperature change. Reaction with the anti-EpCAM antibody HCT116, TUOC1, and TUOS6 cells showed an immune response to the anti-epithelial cellular adhesion molecule (EpCAM) antibody, but not OvBRCA P + 5 cells. Additionally, no anti-EpCAM antibody reaction was observed in the Ovn P + 25, Ov BRCA1 P + 25, or OvBRCA2 P + 25 groups (Fig. 3 ). Chromosomal analysis and mFISH The chromosomal aberrations in the metaphase of each cell line are shown in Table 1 , and there were no differences among SeV- cells. In contrast, among SeV + cells, Ovn cells were mostly normal, whereas Ov BRCA1 and Ov BRCA2 cells showed a high frequency of chromosomal aberrations that increased with passaging. Clonal aberrations were observed in the late phase. Table 1 Chromosomal aberrations in the metaphase of each cell Cell line SeV Passage No. Chromosome No. Aberration Chromosome rearrangement 44 45 46 47 Total ctg/ ictg ctb/ ictb exh Total Ratio Translocation, deletion, dicentric, etc. Total Ratio Ovn normal ー P + 1 0 1 24 0 25 0 0 0 0/25 0% 0/25 0% + P + 5 (early) 0 0 25 0 25 0 0 0 0/25 0% 46,XX,del(5q) 1/25 4.0% + P + 25 (late) 0 1 24 0 25 0 0 0 0/25 0% 46,XX,t(3;16) 46,XX,-22,-22,+2mar 1 2 3/25 12% Ovn BRCA1 ー P + 3 0 1 23 1 25 1 0 0 1/25 4% 47,XX,+mar 1/25 4% + P + 6 (early) 0 1 24 0 25 0 0 0 0/25 0% 46,XX,add(11p),-19,+mar 46,XX,t(6;11) 46,XX,del(7q) 46,XX,-7,-8,+2mar 46,XX,add(5p) 46,XX,del(2p),del(3q) 46,XX,add(1q) 46,XX,del(3p),del(5q),del(7p) 46,XX,del(Xq),add(11q) 1 1 1 1 1 1 1 1 1 9/25 36% + P + 25 (late) 0 0 25 0 25 0 4 0 4/25 16% 46,XX,i(7q),i(8q),add(9q) 46,XX,add(1q),del(1q) 46,XX,i(9q) 45,XX,+1,add(1q),del(1q),-6,-22 1 1 5 1 8/25 32% Ovn BRCA2 ー P + 3 0 0 25 0 25 0 1 0 1/25 4% 0/25 0% + P + 5 (early) 0 0 25 0 25 0 0 0 0/25 0% 46,XX,t(4;15) 46,XX,add(4q) 46,XX,add(14p) 46,XX,del(18p) 46,XX,del(2q) 46,XX,del(17p) 1 2 1 1 1 1 7/25 28% + P + 25 (late) 0 0 25 0 25 0 0 0 0/25 8% 46,XX,t(7;14) 46,XX,dic(1;21) 46,XX,dic(1;8) 46,XX,del(1p) 46,XX,add(14q) 46,XX,add(18q) 1 1 1 1 2 1 7/25 28% Among SeV + cells, Ovn cells were mostly normal, whereas Ov BRCA1 and Ov BRCA2 cells showed a high frequency of chromosomal aberrations that increased with passaging mFISH showed translocations of chromosomes 1 and 22, X and 3, X and 7, or 8 and 9 as well as duplication of chromosome 8 in OvBRCA1 P + 6 and P + 25 and deletion of chromosome 6 in OvBRCA2 P + 4 (Fig. 4 A–D). Human transcriptome sequencing analysis The read lengths and number of assembled transcripts for each cell line are listed in Table 2 . A comparison of the expression levels between samples (Online Resource 4) and the results of the clustering analysis are shown in Fig. 5 . Table 3 presents the list of genes with a high frequency of expression among samples: MMP1 , CCL2 , IGFBP6 , GREM1 , CFB , CXCL1 , CXCL2 , PAPPA , RAB27B , FBLN2 , and C3 in Ov BRCA1 and Ov BRCA2 were highly expressed compared with their expression in Ovn. Table 2 Read lengths and number of assembled transcripts for each cell line in human transcriptome sequencing analysis Sample Direction Original Modified Ratio (%) (modified/original) Read length (bp) No. of reads No. of reads Ovn SeV+ Forward 101 22,265,585 22,249,233 99.93 Reverse 101 22,265,585 22,249,233 99.93 Ovn SeV- Forward 101 22,629,854 22,611,040 99.92 Reverse 101 22,629,854 22,611,040 99.92 OvBRCA1 SeV+ Forward 101 23,333,820 23,319,444 99.94 Reverse 101 23,333,820 23,319,444 99.94 OvBRCA2 SeV+ Forward 101 23,177,522 23,161,169 99.93 Reverse 101 23,177,522 23,161,169 99.93 OvBRCA2 SeV- Forward 101 20,492,659 20,476,948 99.92 Reverse 101 20,492,659 20,476,948 99.92 Table 3 Human transcriptome sequencing analysis Genes with more than 2-fold difference in expression Ovn < Ov BRCA1 Ovn < Ov BRCA2 GREM1 PAMR1 C3 CXCL1 CFB TNC CXCL1 PLAU CXCL2 GREM1 CSF3 CXCL2 SEMA5A C3 VCAM1 SEMA5A PTGS2 PAPPA PAPPA HMCN1 IL6 RAB27B RAB27B CFB CCL5 CPE HMCN1 CHL1 LAMA5 , MIR4758 AADAC FBLN2 FBLN2 CCL2 GBP4 C1R IGFBP6 IGFBP6 SPOCK1 GBP4 IL6 PCOLCE CCL2 TNC NFKBIZ , NXPE3 IGFBP1 MMP1 MMP1 SOX4 CPE ANGPTL2 TIMP3 PRRX1 ANGPTL2 MMP16 CPZ TMEM132A PDK4 OASL SLC7A2 MMP1 , CCL2 , IGFBP6 , GREM1 , CFB , CXCL1 , CXCL2 , PAPPA , RAB27B , FBLN2 , and C3 were highly expressed in Ov BRCA1 or and Ov BRCA2 compared with their levels in Ovn cells Real-time PCR MMP1 , RAB27B , CCL2, PAPPA , GREM1 were highly expressed in Ov BRCA1 compared with their expression in Ovn, whereas MMP1 , CXCL1, RAB27B, CCL2 , PAPPA and GREM1 were significantly highly expressed in Ov BRCA2 (Fig. 6 ). Discussion To the best of our knowledge, this is the first study to successfully establish immortalized normal ovarian epithelial cells and ovarian epithelial cells from patients with germline BRCA1 (g BRCA1 ) or germline BRCA2 (g BRCA2 ) mutations using SeV vectors carrying three immortalization genes. In chromosomal analyses, the presence of g BRCA1 or g BRCA2 mutations suggests that oncogenesis may be in progress because of SeV immortalization. Hence, these cells are useful for analyzing the oncogenic processes in ovarian cancer. Additionally, in the next-generation sequencing analysis, the cell lines with g BRCA1 or g BRCA2 mutation showed significantly high expression of genes such as MMP1 compared to the normal ovarian epithelium. This finding indicates that these genes may be important in oncogenesis. Chromosomal structural abnormalities were observed during the immortalization of ovarian epithelial cells using SeV. Their frequency was higher in Ov BRCA1 and Ov BRCA2 than in Ovn. Possible reasons for this include the instability of germline BRCA mutations and a combination of the effects of SeV infection and passaging. Age-related factors may also be involved, as the age of the germline BRCA1 carrier was 57 years and that of the germline BRCA2 carrier was 53 years. Nonetheless, as age increases, the age of oocytes and frequency of chromosomal abnormalities increase [ 12 ]. Furthermore, ovarian cancer in g BRCA1 mutation carriers has been reported to occur more frequently at a younger age than in g BRCA2 mutation carriers [ 13 – 17 ]. However, as the present study was conducted on a single case, we will discuss the results after accumulating more cases and conducting similar studies in the future. We also examined vascular endothelial cells and stem cells and found no chromosomal abnormalities in these cells, suggesting that the changes may be unique to the ovaries (Oshimura, et al., Submitted). Konishi et al. [ 5 ] established immortalized cells by transfecting human mammary epithelial cells with h-TERT and generated BRCA1 -deficient cells to elucidate the development mechanism of breast cancer. According to their study results, haploinsufficiency of BRCA1 may accelerate the development of hereditary breast cancer by promoting genetic mutations. In that study, instability was induced by the deletion of BRCA in normal cells. However, in the present study, we established immortalized normal ovarian epithelial cells and cells from ovarian epithelial specimens obtained from patients with germline BRCA1 or germline BRCA2 and analyzed their properties. The use of these cell lines may contribute to the elucidation of oncogenic processes, as viral infections of unstable structures are thought to induce further chromosomal aberrations. The transcriptome analysis of each cell line and real-time PCR showed that MMP1 and several other genes were expressed at significantly high levels in the ovarian surface epithelial cells of BRCA1 and BRCA2 carriers. Germline BRCA1 or BRCA2 carriers are at a higher risk of developing ovarian and breast cancers than non-carriers. MMPs are a type of extracellular matrix protein, and Yokoi et al. reported that MMP1 is an indicator of poor prognosis of ovarian cancer. Additionally, extracellular vesicles carrying MMP1 mRNA, which induce the apoptosis of mesothelial cells, were found in the ascites fluid of patients with cancer [ 18 ]. In contrast, a meta-analysis of MMPs as risk factors for ovarian cancer reported no significant association; nonetheless, additional research is warranted [ 19 ]. In the future, we plan to confirm MMP1 expression using frozen tissues and ovarian serous carcinoma cell lines from g BRCA1 or g BRCA2 mutation carriers. In conclusion, the immortalized cell lines established in this study have the potential to elucidate a new oncogenic process by confirming the gene expression in normal and abnormal clones of the same cell line and via epigenomic analysis. This study has several strengths. First, SeV functions as an RNA in the cytoplasm and is not incorporated into the chromosomes of target cells, thus eliminating genotoxicity. Second, efficient gene transfer can be achieved in a short time with low infection titers or target cells. Third, transgenic cells do not produce infectious viral particles; hence, they can be safely used. Owing to these characteristics, SeV vectors are being developed as novel gene delivery systems, called cytoplasmic RNA vectors, for gene therapy and gene vaccines. They are being increasingly used as research tools in biotechnology and for developing bioproducts. In this study, we established immortalized cells using this method. This study also has some limitations. First, this study was based on the analysis of one case each, and it was not possible to fully examine whether the chromosomal abnormalities were unique to the case or ovarian cancer. However, the absence of abnormalities in other immortalized cell lines, such as vascular endothelial cells, suggests that these changes may be unique to ovarian cells. SeV vectors exist in the cytoplasm in the form of RNA throughout their life cycle and are not susceptible to insertional mutations or structural changes in chromosomes because of incorporation into the chromosomal genome. This is in contrast to retroviral vectors, which express genes by incorporation into chromosomes, or adenovirus vectors, adeno-associated virus vectors, and plasmid vectors, which exist as DNA in the cell nucleus away from chromosomes and can be incorporated into chromosomes at a certain frequency. Second, the SeV vector is temperature-sensitive, and temperature changes can introduce mutations at the amino acid level to facilitate the removal of the vector. Hence, the CO 2 incubator temperature must be set at 35°C. Incubating cells at 37°C may lead to changes in cell characteristics and senescence. Therefore, it is essential to handle these cells with utmost care and culture them in a CO 2 incubator with temperature control. Moreover, immunocytochemical staining yielded positive results for keratin in Ovn Passage + 8, Ov BRCA1 Passage + 9, and Ov BRCA2 Passage + 9 cells and for vimentin in some cells (Online Resource 5). One possibility is that epithelial–mesenchymal transition occurred with cell passage. Another explanation is that some stromal cells may have strayed into the tumor cells. However, when we collect cells from patient specimens, we appropriately sample from the ovarian surface layer and usually confirm that they are epithelial cells. In this study, normal ovarian cells were successfully established using SeV, and ovarian epithelial cells were obtained from patients harboring germline BRCA1 or BRCA2 mutations. These cell lines can be used in research on ovarian tumors in the future, to understand the mechanism of ovarian cancer development, as well as in research to identify drugs and new diagnostic markers. Additionally, our results suggest that MMP1 might be important during the development of ovarian cancer with BRCA mutations. Although the establishment of immortalized cells of the normal ovarian epithelium has been previously reported, we maintained ovarian epithelial cells generated from patients with g BRCA1 and g BRCA2 mutations in addition to normal ovarian epithelial cells. Furthermore, our original immortalization technology is safe and easy to use, which is a major advantage. Although some issues remain, this immortalization technology has the potential for further development and it is expected that its application will extend beyond cell lines to include other than ovarian cells. We plan to construct a library of immortalized cell lines in the future and establish a sustainable research infrastructure. Declarations Funding This work was supported by a research grant from the Tottori University Hospital and a Grant-in-Aid for Scientific Research (Grant number 21K16770) in 2021. Compliance with Ethical Standards Disclosure of potential conflicts of interest The authors, except Mitsuo Oshimura, declare that they have no financial or non-financial interests to disclose. Research involving Human Participants Ethics Statement The study was approved by the Ethics Committee of Tottori University (IRB number: 20A179). Informed consent Consent to participate Informed consent was obtained from all individual participants included in the study. Consent to publish Not applicable. Author Contributions Hiroaki Komatsu, Yasuhiro Kazuki, and Masayo Okawa conceived the study. Hiroaki Komatsu and Masayo Okawa developed the statistical analysis plan and conducted statistical analyses. All authors contributed to the interpretation of the results. Hiroaki Komatsu drafted the original manuscript. Yasuhiro Kazuki and Masayo Okawa supervised the conduct of this study. All authors reviewed the manuscript draft and revised it critically for intellectual content. All authors approved the final version of the manuscript to be published. Data Availability The authors confirm that the data supporting the findings of this study are available within the article or its supplementary materials. Acknowledgments The authors are grateful to Mitsuhiko Ozaki, Ph.D., from the Department of Experimental Pathology, Department of Life Sciences, Faculty of Medicine, Tottori University, for providing guidance on the research methods. This work was supported by a research grant from the Tottori University Hospital and a Grant-in-Aid for Scientific Research (21K16770) in 2021. References Kurman RJ, Shih LM. The origin and pathogenesis of epithelial ovarian cancer: a proposed unifying theory. Am J Surg Pathol. 2010;34:433–43. Mogos RA, Popovici R, Tanase AE, et al. New approaches in ovarian cancer based on genetics and carcinogenesis hypotheses (Review). Exp Ther Med. 2022;23:423. Cancer Genome Atlas Research Network. Integrated genomic analyses of ovarian carcinoma. Nature. 2011;474:609–15. Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature. 2012;490:61–70. Konishi H, Mohseni M, Tamaki A, et al. Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells. Proc Natl Acad Sci U S A. 2011;108:17773–8. Ohira M, Kikuchi E, Mizuta S, et al. 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Cancer risks for BRCA1 and BRCA2 mutation carriers: results from prospective analysis of EMBRACE. J Natl Cancer Inst. 2013;105:812–22. Yokoi A, Yoshioka Y, Yamamoto Y, et al. Malignant extracellular vesicles carrying MMP1 mRNA facilitate peritoneal dissemination in ovarian cancer. Nat Commun. 2017;8:14470. Zhu XM, Sun WF. Association between matrix metalloproteinases polymorphisms and ovarian cancer risk: A meta-analysis and systematic review. PLoS ONE. 2017;12:e0185456. Tables Table 1. Chromosomal aberrations in the metaphase of each cell Cell line SeV Passage No. Chromosome No. Aberration Chromosome rearrangement 44 45 46 47 Total ctg/ ictg ctb/ ictb exh Total Ratio Translocation, deletion, dicentric, etc. Total Ratio Ovn normal ー P+1 0 1 24 0 25 0 0 0 0/25 0% 0/25 0% + P+5 (early) 0 0 25 0 25 0 0 0 0/25 0% 46,XX,del(5q) 1/25 4.0% + P+25 (late) 0 1 24 0 25 0 0 0 0/25 0% 46,XX,t(3;16) 46,XX,-22,-22,+2mar 1 2 3/25 12% Ovn BRCA1 ー P+3 0 1 23 1 25 1 0 0 1/25 4% 47,XX,+mar 1/25 4% + P+6 (early) 0 1 24 0 25 0 0 0 0/25 0% 46,XX,add(11p),-19,+mar 46,XX,t(6;11) 46,XX,del(7q) 46,XX,-7,-8,+2mar 46,XX,add(5p) 46,XX,del(2p),del(3q) 46,XX,add(1q) 46,XX,del(3p),del(5q),del(7p) 46,XX,del(Xq),add(11q) 1 1 1 1 1 1 1 1 1 9/25 36% Table 2. Read lengths and number of assembled transcripts for each cell line in human transcriptome sequencing analysis Sample Direction Original Modified Ratio (%) (modified/original) Read length (bp) No. of reads No. of reads Ovn SeV+ Forward 101 22,265,585 22,249,233 99.93 Reverse 101 22,265,585 22,249,233 99.93 Ovn SeV- Forward 101 22,629,854 22,611,040 99.92 Reverse 101 22,629,854 22,611,040 99.92 OvBRCA1 SeV+ Forward 101 23,333,820 23,319,444 99.94 Reverse 101 23,333,820 23,319,444 99.94 OvBRCA2 SeV+ Forward 101 23,177,522 23,161,169 99.93 Reverse 101 23,177,522 23,161,169 99.93 OvBRCA2 SeV- Forward 101 20,492,659 20,476,948 99.92 Reverse 101 20,492,659 20,476,948 99.92 Table 3 Human transcriptome sequencing analysis Genes with more than 2-fold difference in expression Ovn < Ov BRCA1 Ovn < Ov BRCA2 GREM1 PAMR1 C3 CXCL1 CFB TNC CXCL1 PLAU CXCL2 GREM1 CSF3 CXCL2 SEMA5A C3 VCAM1 SEMA5A PTGS2 PAPPA PAPPA HMCN1 IL6 RAB27B RAB27B CFB CCL5 CPE HMCN1 CHL1 LAMA5 , MIR4758 AADAC FBLN2 FBLN2 CCL2 GBP4 C1R IGFBP6 IGFBP6 SPOCK1 GBP4 IL6 PCOLCE CCL2 TNC NFKBIZ , NXPE3 IGFBP1 MMP1 MMP1 SOX4 CPE ANGPTL2 TIMP3 PRRX1 ANGPTL2 MMP16 CPZ TMEM132A PDK4 OASL SLC7A2 MMP1 , CCL2 , IGFBP6 , GREM1 , CFB , CXCL1 , CXCL2 , PAPPA , RAB27B , FBLN2 , and C3 were highly expressed in Ov BRCA1 or and Ov BRCA2 compared with their levels in Ovn cells Supplementary Files Supportinginformation.docx Cite Share Download PDF Status: Published Journal Publication published 14 Apr, 2024 Read the published version in Human Cell → Version 1 posted Reviewers agreed at journal 07 Mar, 2024 Reviewers invited by journal 07 Mar, 2024 Editor assigned by journal 04 Mar, 2024 First submitted to journal 01 Mar, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-4012283\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":276951067,\"identity\":\"dc2db8db-6fd8-4da6-a5e0-ee8034969294\",\"order_by\":0,\"name\":\"Hiroaki Komatsu\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA80lEQVRIie3PPQrCMBiA4ZSAiw2uKQXPUCcVj+Lk5JS9g0qmOqi7gz9XUAJxTQm0i8VVcPEACoJLwQ62FcWlBjfBvMOXDN9DCAA63Q+GnxcnG6abDgipmogX2WbE+IYgL5sKYg1HneslkaAOyp3jedFvV4YpiV1eSGwzYtj3JGhSxGpzHpKpNKgx3h4KSRWTFfapBI5A3EY8IDQl0PA+EhaLJCebG5oFZKkiNiYci9LjFYhoj6xUxBpHvBF5XdORiFmzQJB1SvxPf8HhhO3dpFV1wsn6cuoNyGIn/WPsFpNnJoD5KfMplPtvDb5Z1ul0uj/pDhqxXngOJW+UAAAAAElFTkSuQmCC\",\"orcid\":\"https://orcid.org/0000-0002-4507-6848\",\"institution\":\"Tottori University Hospital: Tottori Daigaku Igakubu Fuzoku Byoin\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Hiroaki\",\"middleName\":\"\",\"lastName\":\"Komatsu\",\"suffix\":\"\"},{\"id\":276951068,\"identity\":\"39cd7fd7-98fa-4885-bd0d-029c781665f2\",\"order_by\":1,\"name\":\"Masayo okawa\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University Hospital: Tottori Daigaku Igakubu Fuzoku Byoin\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Masayo\",\"middleName\":\"\",\"lastName\":\"okawa\",\"suffix\":\"\"},{\"id\":276951069,\"identity\":\"da125cac-cc84-4dff-bb7f-403a72cb9c2e\",\"order_by\":2,\"name\":\"Yasuhiro Kazuki\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yasuhiro\",\"middleName\":\"\",\"lastName\":\"Kazuki\",\"suffix\":\"\"},{\"id\":276951070,\"identity\":\"d534624c-c25d-4b5d-b253-175cbbc91e33\",\"order_by\":3,\"name\":\"kanako Kazuki\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"kanako\",\"middleName\":\"\",\"lastName\":\"Kazuki\",\"suffix\":\"\"},{\"id\":276951071,\"identity\":\"95f2fedb-0be4-4df9-a941-052928f92efe\",\"order_by\":4,\"name\":\"Genki Hichiwa\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Genki\",\"middleName\":\"\",\"lastName\":\"Hichiwa\",\"suffix\":\"\"},{\"id\":276951072,\"identity\":\"f28ba52e-c192-4d08-803b-d204126e4f3e\",\"order_by\":5,\"name\":\"kazuhito Shimoya\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"kazuhito\",\"middleName\":\"\",\"lastName\":\"Shimoya\",\"suffix\":\"\"},{\"id\":276951073,\"identity\":\"93f78a27-805b-4413-9dba-82e0a8e02225\",\"order_by\":6,\"name\":\"Shin Sato\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Shin\",\"middleName\":\"\",\"lastName\":\"Sato\",\"suffix\":\"\"},{\"id\":276951074,\"identity\":\"4727e8c1-1978-41bb-862f-13498c1c28e7\",\"order_by\":7,\"name\":\"Fuminori Taniguchi\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Fuminori\",\"middleName\":\"\",\"lastName\":\"Taniguchi\",\"suffix\":\"\"},{\"id\":276951075,\"identity\":\"3aff289d-bf5e-4f6f-ac49-c8a5b2b2e275\",\"order_by\":8,\"name\":\"Mitsu Oshimura\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Transchromosomics Inc\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Mitsu\",\"middleName\":\"\",\"lastName\":\"Oshimura\",\"suffix\":\"\"},{\"id\":276951076,\"identity\":\"4fef3dbd-9522-4596-b73c-34dafabd365a\",\"order_by\":9,\"name\":\"Tasuku Harada\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Tottori University: Tottori Daigaku\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Tasuku\",\"middleName\":\"\",\"lastName\":\"Harada\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-03-04 15:16:41\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4012283/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4012283/v1\",\"draftVersion\":[],\"editorialEvents\":[{\"content\":\"https://doi.org/10.1007/s13577-024-01064-z\",\"type\":\"published\",\"date\":\"2024-04-14T14:49:07+00:00\"}],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":52451324,\"identity\":\"babbfcf8-6bc0-41fa-97cc-efbc2fb6a507\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:12:33\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":655079,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eAll-in-one microscopy of Ovn, OvBRCA1, and OvBRCA2 cell lines\\u003c/p\\u003e\\n\\u003cp\\u003eGFP and OFP fluorescence at the same magnification as in brightfield microscopy\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/e857c71e1d4da25e53407601.png\"},{\"id\":52451320,\"identity\":\"331b33c5-072b-4a8e-896f-df81e1569db5\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:12:33\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":588723,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eGrowth curves of Ovn, OvBRCA1, and OvBRCA2 cells\\u003c/p\\u003e\\n\\u003cp\\u003eSeV+ cells continue to proliferate, whereas SeV- cells do not proliferate early\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/c08e82ff945c6cda4e70ecc4.png\"},{\"id\":52451322,\"identity\":\"353f2876-1538-4379-9b6e-c1092836e85c\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:12:33\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":186056,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eAnti-EpCAM antibody reaction observed via flow cytometry of each cell\\u003c/p\\u003e\\n\\u003cp\\u003eThe colon cancer cell line HCT-116 and epithelial ovarian cancer cell lines TU-OC-1 and TU-OS-6 showed a response, whereas the immortalized cell lines did not show an antibody response either during early infection or in long-term culture\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/f995e4231b519e964c4b595b.png\"},{\"id\":52451323,\"identity\":\"b9336e9c-6439-4e7a-8032-0b476a069ad7\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:12:33\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":324569,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eRepresentative chromosomal structural aberrations, as revealed via mFISH\\u003c/p\\u003e\\n\\u003cp\\u003eKaryotype notation does not conform to ISCN\\u003c/p\\u003e\\n\\u003cp\\u003eThe white arrow indicates chromosome rearrangement. Scale bar: 10 μm\\u003c/p\\u003e\\n\\u003cp\\u003eA) Ovn BRCA1 SeV- P+3, 46,XX, normal\\u003c/p\\u003e\\n\\u003cp\\u003eB) Ovn BRCA1 SeV+ P+6, 45,XX,del(Xq),der(1;22),der(3)t(X;3),chtg(7),der(7)t(X;7)\\u003c/p\\u003e\\n\\u003cp\\u003eC) Ovn BRCA1 SeV+ P+25, 46,XX,dup(8),der(9)t(8;9)\\u003c/p\\u003e\\n\\u003cp\\u003eD) Ovn BRCA2 SeV+ P+4, 46,XX,del(6p)\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/4154ca620ed10096d89bfe4e.png\"},{\"id\":52453256,\"identity\":\"0173f296-5e9f-4252-ad4c-819aeb5bcb2f\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:20:36\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1589813,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eClustering analysis in human transcriptome sequencing analysis\\u003c/p\\u003e\\n\\u003cp\\u003eThe results of the cluster analysis of Ovn with OvBRCA1 and OvBRCA2 are shown. Each analysis was classified into one of five categories. The most characteristic feature was that the slope of cluster 4 was different\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/080e6d77899a8e591a7ecaba.png\"},{\"id\":52453257,\"identity\":\"dfc7262a-d398-469d-9040-cea66a15e768\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:20:36\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":117940,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eReal-time PCR\\u003c/p\\u003e\\n\\u003cp\\u003eMMP1, RAB27B, and PAPPA were highly expressed in Ov BRCA1 compared with those in Ovn, whereas MMP1, CCL2, GREM1, CXCL1, and PAPPA were significantly highly expressed in Ov BRCA2\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/f25ca90c40a4d71432873fba.png\"},{\"id\":54650856,\"identity\":\"bb37a851-e648-43e4-a83a-25dd466217b0\",\"added_by\":\"auto\",\"created_at\":\"2024-04-14 14:49:16\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":1898567,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/64ab5a71-0477-4d6d-a5ba-7aa3c1fd0ea3.pdf\"},{\"id\":52451326,\"identity\":\"15661dcb-2896-4b9b-92f2-e3e06e08ce61\",\"added_by\":\"auto\",\"created_at\":\"2024-03-11 19:12:34\",\"extension\":\"docx\",\"order_by\":15,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":234345,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"Supportinginformation.docx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4012283/v1/7aa65c33da63f3a9e069a68a.docx\"}],\"financialInterests\":\"\",\"formattedTitle\":\"Characterization of immortalized ovarian epithelial cells with BRCA1/2 mutation\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eThe pathogenesis of ovarian cancer is becoming clearer in terms of the histological and clinicopathological background. A recent study has shown that the origin of ovarian cancer, especially high-grade serous carcinoma, is serous tubal intraepithelial carcinoma [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e]. However, analysis at the molecular level remains insufficient [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e], which may be attributed to the lack of sufficient cell lines derived from normal ovarian epithelium, benign lesions, and borderline malignant lesions to allow for a detailed analysis of the mechanisms involved in malignant transformation. \\u003cem\\u003eBRCA1\\u003c/em\\u003e and \\u003cem\\u003eBRCA2\\u003c/em\\u003e are crucial genes involved in ovarian cancer development, and germline mutations in these genes have been linked to a high frequency of breast and ovarian cancers [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. However, there are no reports on predictors or appropriate diagnostic markers for cancer development in patients with \\u003cem\\u003eBRCA1\\u003c/em\\u003e and \\u003cem\\u003eBRCA2\\u003c/em\\u003e mutations, making regular examination and measurement of tumor markers crucial. Although \\u003cem\\u003eBRC\\u003c/em\\u003eA mutations in mammary epithelial cells have been related to genetic instability, studies on \\u003cem\\u003eBRCA\\u003c/em\\u003e in immortalized cells are intriguing [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eEstablishing normal ovarian epithelial cells and cells with \\u003cem\\u003eBRCA1\\u003c/em\\u003e/2 mutations will considerably expand the scope of research, including the analysis of genes involved in carcinogenesis, factors that inhibit carcinogenesis, and mechanisms underlying carcinogenesis. However, ovarian epithelial cells cannot be passaged over a long period and must be collected and cultured each time as research material. This process is difficult because ovaries are intraperitoneal organs.\\u003c/p\\u003e \\u003cp\\u003eSendai virus (SeV) vectors have a genome of single-stranded minus-strand RNA that can be introduced into non-dividing and dividing cells and have a high ability to express foreign genes [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e]. As the vector genome remains in the cytoplasm, it does not affect host chromosomes and does not induce structural changes or mutations; therefore, it has been used as a tool to identify the causes of rare diseases and devise suitable treatments [\\u003cspan additionalcitationids=\\\"CR8 CR9\\\" citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e]. In the present study, we attempted to establish immortalized normal ovarian epithelial cells and ovarian epithelial cells from patients with germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e or \\u003cem\\u003eBRCA2\\u003c/em\\u003e mutations using SeV vectors and to confirm whether the properties of each cell line could be analyzed.\\u003c/p\\u003e\"},{\"header\":\"Materials and Methods\",\"content\":\"\\u003cp\\u003e \\u003cem\\u003ePatient background\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003ePatient A was 49 years old and underwent total hysterectomy and bilateral ovarian and fallopian tube resection for atypical endometrial hyperplasia; the epithelium of the ovarian surface layer was extracted via a fine surgical cut using a scalpel. Patient B, aged 57 years, developed breast cancer, and genetic testing revealed a \\u003cem\\u003eBRCA1\\u003c/em\\u003e mutation (C.405_406 del (P.Arg 136 Thrfs*5)) and was diagnosed with hereditary breast/ovarian cancer (HBOC). Patient C developed breast cancer at the age of 53 years, and genetic testing revealed a \\u003cem\\u003eBRCA2\\u003c/em\\u003e mutation (C.1813 del (P.lle 605 Thrfs*9)) and was diagnosed with HBOC. In Patients B and C, the ovarian epithelium was collected via abrasion from surgical specimens using risk-reducing salpingo-oophorectomy (RRSO). In none of these cases of ovarian or fallopian tube carcinoma, the serous tubal intraepithelial carcinoma (STIC) was grossly or histopathologically observed in hematoxylin and eosin (HE)-stained specimens with paraffin block embedding.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eEstablishment of primary cell lines\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe surface of the excised ovarian tissue was cut into small pieces with a scalpel, and 10 mL of 1% collagenase\\u0026thinsp;+\\u0026thinsp;Dulbecco\\u0026rsquo;s modified Eagle\\u0026rsquo;s medium (DMEM)/Ham\\u0026rsquo;s F-12 with \\u003cspan type=\\\"SmallCaps\\\" class=\\\"SmallCaps\\\" name=\\\"Emphasis\\\"\\u003el\\u003c/span\\u003e-glutamine and sodium pyruvate (Product Number: 045-30665) was added to a 15 mL tube. The tube was incubated horizontally in a 37\\u0026deg;C incubator (100 rpm) for approximately 24 h. Subsequently, the mixture was filtered twice into a 50 mL tube using a 70 \\u0026micro;m strainer (Product Number: 352350). The mixture was then centrifuged at 1,300 rpm for 8 min, and the supernatant was discarded. The cells were washed with 2 mL of DMEM/Ham\\u0026rsquo;s F12, and then 8 mL of DMEM/F12 was added; the mixture was transferred to a 15 mL tube and filtered into a 50 mL tube using a 40 \\u0026micro;m (Product Number: 352340) strainer. The cells were seeded in a 60 mm dish with DMEM/Ham\\u0026rsquo;s F12\\u0026thinsp;+\\u0026thinsp;20% fetal bovine serum (FBS)\\u0026thinsp;+\\u0026thinsp;1.0 penicillin\\u0026ndash;streptomycin and incubated in a CO\\u003csub\\u003e2\\u003c/sub\\u003e incubator at 37\\u0026deg;C. The next day, blood cells were removed by washing with phosphate-buffered saline (PBS), and the cells were seeded at a density of 2.0 \\u0026times; 10\\u003csup\\u003e5\\u003c/sup\\u003e cells/well in a 24-well plate. On the following day, the cells were infected with an SeV vector carrying three immortalization genes (\\u003cem\\u003ehTERT\\u003c/em\\u003e, \\u003cem\\u003eBmi1\\u003c/em\\u003e, and \\u003cem\\u003eSV40T\\u003c/em\\u003e). The SeV vectors were used for gene loading and vector production by ID Pharma, Inc. Finally, the infected cells were subjected to long-term passaging.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eSeV vector infection\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe cells were seeded (2 \\u0026times; 10\\u003csup\\u003e5\\u003c/sup\\u003e cells) in 24-well plates. The next day, the cells were infected with SeV-hTERT (with simultaneous enhanced green fluorescent protein (EGFP) loading), SeV-Bmi1 (with simultaneous orange fluorescent protein (OFP) loading), and SeV-SV40T in a safety cabinet (the amount of virus used for infection is not disclosed owing to a pending patent).\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eFlow cytometry\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe cell count was 2.5 \\u0026times; 10\\u003csup\\u003e5\\u003c/sup\\u003e cells/100 mL PBS\\u0026thinsp;+\\u0026thinsp;1% FBS/well. Immortalized cells derived from human tissues (Ovn Passage\\u0026thinsp;+\\u0026thinsp;20, Ov BRCA1 Passage\\u0026thinsp;+\\u0026thinsp;5, Ov BRCA1 Passage\\u0026thinsp;+\\u0026thinsp;20, Ov BRCA2 Passage\\u0026thinsp;+\\u0026thinsp;20) were used along with positive control cells (colon cancer cell line HCT116, ovarian clear cell carcinoma cell line TU-OC-1 [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e], and ovarian serous adenocarcinoma cell line TU-OS-6). Flow cytometry was performed with accutase treatment for 1\\u0026ndash;3 min when the confluency of all cell lines reached approximately 70% (Online Resource 1). The survival rate was approximately 90% for all cell lines.\\u003c/p\\u003e \\u003cp\\u003eThe primary antibodies (1 mg/100 mL/well) used were anti-EpCAM antibody (hZAK3A101; original stock: 1.105 mg/mL; 20210505) and negative control antibody (hIgG; original stock: 1 mg/mL; Invitrogen; 12000C; S1257958). The cells were incubated at 4\\u0026deg;C for 1 h and then washed twice with PBS\\u0026thinsp;+\\u0026thinsp;1% FBS (200 mL/well). The secondary antibody anti-hIgG antibody Alexa 488 (original stock: 1.5 mg/mL; Jackson ImmunoResearch, 109-545-190) was added (0.075 mg/30 mL/well) to the cells, and the mixture was incubated at 4\\u0026deg;C for 1 h and then washed twice with PBS\\u0026thinsp;+\\u0026thinsp;1% FBS (200 mL/well).\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eCloning of ovarian cells\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe established cell lines were seeded at low concentrations in a 100 mm dish. The cells were observed for more than 40 days, and the area where colonies formed from single cells was observed using an all-in-one microscope to confirm that both green fluorescent protein (GFP) and OFP were detected. The cells were selected under a microscope and seeded in 96-well plates. This step was repeated multiple times to pass on more than 50 cloned cells, scaling up from 96-well plates to 24-well plates, 12-well plates, 6-well plates, 60 mm dishes, and 100 mm dishes, in that order. Cells that could be passed on were designated as cloning cells. The cloned cells were defined as those that could be passaged.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eChromosome analysis\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eEach cell line was seeded in a 60 mm dish, treated synchronously with MAS (Genial Genetic Solutions), and then fixed using the Carnoy\\u0026rsquo;s fixation method. The fixed cells were spread on slides and stained with quinacrine mustard (QH; Sigma\\u0026ndash;Aldrich, Hoechst 33258; Sigma\\u0026ndash;Aldrich) for multicolor fluorescence in situ hybridization (mFISH) (24XCyte: MetaSystems Inc., Altlu\\u0026szlig;heim, Germany), according to the manufacturer\\u0026rsquo;s instructions (MetaSystems). Metaphase images were captured with an AxioImagerZ2 fluorescence microscope (Carl Zeiss GmbH, Jena, Germany) and analyzed using the Ikaros software program (MetaSystems) and ISIS software program. Chromosome karyotyping of metaphase was performed to determine the chromosome number and karyotypic structural aberrations.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eHuman transcriptome sequencing analysis\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eTotal RNA was extracted using the RNeasy Mini Kit (Qiagen; cat. No. 74104, 74106), and quality control was performed using the Agilent 2200 TapeStation to confirm that there was at least 2.0 \\u0026micro;g of RNA. Analysis was performed using NovaSeq 6000 (Illumina). Libraries were prepared using the TruSeq stranded mRNA Library Prep Kit with a library length of 332\\u0026ndash;346 bp. The read length was 101 bp and was analyzed using the multiplex method. Cluster analysis was performed using the k-means method by classifying clusters into five categories according to the slope of the expression variation ratio.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eReal-time polymerase chain reaction\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eTotal RNA was extracted from each cell line using the RNeasy Mini Kit (Qiagen; Cat. Nos. 74104 and 74106). DNase treatment was performed using deoxyribonuclease (RT Grade, Cat. No. 313\\u0026ndash;03161) and the RNaseOUT\\u0026trade; Recombinant Ribonuclease Inhibitor (Cat. No. 10777019).\\u003c/p\\u003e \\u003cp\\u003eA high-capacity cDNA Reverse Transcription Kit (Applied Biosystems; Cat. No. 4368814) was used for cDNA synthesis. Real-time polymerase chain reaction (PCR) was performed using the StepOnePlus Real-Time PCR system (Applied Biosystems) and TaqMan Fast Advanced Master Mix (Applied Biosystems) according to the manufacturer\\u0026rsquo;s protocol.\\u003c/p\\u003e \\u003cp\\u003eTranscript levels were normalized to those of beta-actin (ACTB). The PCR conditions were as follows: 50\\u0026deg;C for 2 min, 95\\u0026deg;C for 20 s, followed by 40 cycles at 95\\u0026deg;C for 1 s and 60\\u0026deg;C for 20 s. Each reaction was performed in triplicate. The PCR primer sequences are shown in Online Resource 2. PCR primers that showed more than 2-fold gene expression in Ov BRCA1 and Ov BRCA2 compared with that in Ovn upon human transcriptome sequencing analysis were selected.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eEthics statement\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eAll patients provided written informed consent following the institutional guidelines. All methods were carried out in accordance with relevant guidelines and regulations. The study was approved by the Ethics Committee of Tottori University (IRB number: 20A179).\\u003c/p\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003e \\u003cem\\u003eMorphological and growth characteristics\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eOvn, Ov BRCA1, and Ov BRCA2 exhibited spindle-shaped or similar round cell morphology. All cell lines showed GFP and OFP fluorescence (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). The growth curves for each cell type are shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. Each SeV-infected cell line showed growth, and SeV-uninfected Ovn, Ov BRCA1, and Ov BRCA2 cells survived for up to 10, 20, and 35 days, respectively; however, no further growth was observed.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eTemperature dependence\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eFor Ov BRCA1, the temperature was increased from 35\\u0026deg;C to 37\\u0026deg;C in the CO\\u003csub\\u003e2\\u003c/sub\\u003e incubator. The intensity of GFP and OFP staining decreased markedly on day 10 after the temperature change (Online Resource 3), and the cells showed no growth after 20 days of temperature change.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eReaction with the anti-EpCAM antibody\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eHCT116, TUOC1, and TUOS6 cells showed an immune response to the anti-epithelial cellular adhesion molecule (EpCAM) antibody, but not OvBRCA P\\u0026thinsp;+\\u0026thinsp;5 cells. Additionally, no anti-EpCAM antibody reaction was observed in the Ovn P\\u0026thinsp;+\\u0026thinsp;25, Ov BRCA1 P\\u0026thinsp;+\\u0026thinsp;25, or OvBRCA2 P\\u0026thinsp;+\\u0026thinsp;25 groups (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eChromosomal analysis and mFISH\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe chromosomal aberrations in the metaphase of each cell line are shown in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e, and there were no differences among SeV- cells. In contrast, among SeV\\u0026thinsp;+\\u0026thinsp;cells, Ovn cells were mostly normal, whereas Ov BRCA1 and Ov BRCA2 cells showed a high frequency of chromosomal aberrations that increased with passaging. Clonal aberrations were observed in the late phase.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eChromosomal aberrations in the metaphase of each cell\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"16\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c8\\\" colnum=\\\"8\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c9\\\" colnum=\\\"9\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c10\\\" colnum=\\\"10\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c11\\\" colnum=\\\"11\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c12\\\" colnum=\\\"12\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c13\\\" colnum=\\\"13\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c14\\\" colnum=\\\"14\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c15\\\" colnum=\\\"15\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c16\\\" colnum=\\\"16\\\"\\u003e\\u003c/div\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eCell line\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eSeV\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003ePassage\\u003c/p\\u003e \\u003cp\\u003eNo.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c8\\\" namest=\\\"c4\\\"\\u003e \\u003cp\\u003eChromosome No.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c13\\\" namest=\\\"c9\\\"\\u003e \\u003cp\\u003eAberration\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c16\\\" namest=\\\"c14\\\"\\u003e \\u003cp\\u003eChromosome rearrangement\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e44\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e45\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e46\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e47\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003eTotal\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003ectg/\\u003c/p\\u003e \\u003cp\\u003eictg\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003ectb/\\u003c/p\\u003e \\u003cp\\u003eictb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003eexh\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003eTotal\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003eRatio\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003eTranslocation, deletion, dicentric, etc.\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003eTotal\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003eRatio\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"2\\\" rowspan=\\\"3\\\"\\u003e \\u003cp\\u003eOvn normal\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eー\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e24\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;5\\u003c/p\\u003e \\u003cp\\u003e(early)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e46,XX,del(5q)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e4.0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;25\\u003c/p\\u003e \\u003cp\\u003e(late)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e24\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e46,XX,t(3;16)\\u003c/p\\u003e \\u003cp\\u003e46,XX,-22,-22,+2mar\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003cp\\u003e3/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e12%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"2\\\" rowspan=\\\"3\\\"\\u003e \\u003cp\\u003eOvn BRCA1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eー\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e23\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e1/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e4%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e47,XX,+mar\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e4%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;6\\u003c/p\\u003e \\u003cp\\u003e(early)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e24\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e46,XX,add(11p),-19,+mar\\u003c/p\\u003e \\u003cp\\u003e46,XX,t(6;11)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(7q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,-7,-8,+2mar\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(5p)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(2p),del(3q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(1q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(3p),del(5q),del(7p)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(Xq),add(11q)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e9/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e36%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;25\\u003c/p\\u003e \\u003cp\\u003e(late)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e4/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e16%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e46,XX,i(7q),i(8q),add(9q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(1q),del(1q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,i(9q)\\u003c/p\\u003e \\u003cp\\u003e45,XX,+1,add(1q),del(1q),-6,-22\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e5\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e8/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e32%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"2\\\" rowspan=\\\"3\\\"\\u003e \\u003cp\\u003eOvn BRCA2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eー\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e1/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e4%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;5\\u003c/p\\u003e \\u003cp\\u003e(early)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e0%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e46,XX,t(4;15)\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(4q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(14p)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(18p)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(2q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(17p)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e7/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e28%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eP\\u0026thinsp;+\\u0026thinsp;25\\u003c/p\\u003e \\u003cp\\u003e(late)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c8\\\"\\u003e \\u003cp\\u003e25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c9\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c10\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c11\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c12\\\"\\u003e \\u003cp\\u003e0/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c13\\\"\\u003e \\u003cp\\u003e8%\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c14\\\"\\u003e \\u003cp\\u003e46,XX,t(7;14)\\u003c/p\\u003e \\u003cp\\u003e46,XX,dic(1;21)\\u003c/p\\u003e \\u003cp\\u003e46,XX,dic(1;8)\\u003c/p\\u003e \\u003cp\\u003e46,XX,del(1p)\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(14q)\\u003c/p\\u003e \\u003cp\\u003e46,XX,add(18q)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c15\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e2\\u003c/p\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003cp\\u003e7/25\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c16\\\"\\u003e \\u003cp\\u003e28%\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"16\\\"\\u003eAmong SeV\\u0026thinsp;+\\u0026thinsp;cells, Ovn cells were mostly normal, whereas Ov BRCA1 and Ov BRCA2 cells showed a high frequency of chromosomal aberrations that increased with passaging\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003emFISH showed translocations of chromosomes 1 and 22, X and 3, X and 7, or 8 and 9 as well as duplication of chromosome 8 in OvBRCA1 P\\u0026thinsp;+\\u0026thinsp;6 and P\\u0026thinsp;+\\u0026thinsp;25 and deletion of chromosome 6 in OvBRCA2 P\\u0026thinsp;+\\u0026thinsp;4 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003eA\\u0026ndash;D).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eHuman transcriptome sequencing analysis\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe read lengths and number of assembled transcripts for each cell line are listed in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. A comparison of the expression levels between samples (Online Resource 4) and the results of the clustering analysis are shown in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e. Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e presents the list of genes with a high frequency of expression among samples: \\u003cem\\u003eMMP1\\u003c/em\\u003e, \\u003cem\\u003eCCL2\\u003c/em\\u003e, \\u003cem\\u003eIGFBP6\\u003c/em\\u003e, \\u003cem\\u003eGREM1\\u003c/em\\u003e, \\u003cem\\u003eCFB\\u003c/em\\u003e, \\u003cem\\u003eCXCL1\\u003c/em\\u003e, \\u003cem\\u003eCXCL2\\u003c/em\\u003e, \\u003cem\\u003ePAPPA\\u003c/em\\u003e, \\u003cem\\u003eRAB27B\\u003c/em\\u003e, \\u003cem\\u003eFBLN2\\u003c/em\\u003e, and \\u003cem\\u003eC3\\u003c/em\\u003e in Ov BRCA1 and Ov BRCA2 were highly expressed compared with their expression in Ovn.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eRead lengths and number of assembled transcripts for each cell line in human transcriptome sequencing analysis\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"6\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eSample\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eDirection\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eOriginal\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eModified\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eRatio (%) (modified/original)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eRead length (bp)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eNo. of reads\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eNo. of reads\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eOvn SeV+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eForward\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e22,265,585\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e22,249,233\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.93\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eReverse\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e22,265,585\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e22,249,233\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.93\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eOvn SeV-\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eForward\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e22,629,854\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e22,611,040\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.92\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eReverse\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e22,629,854\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e22,611,040\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.92\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eOvBRCA1 SeV+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eForward\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e23,333,820\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e23,319,444\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.94\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eReverse\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e23,333,820\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e23,319,444\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.94\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eOvBRCA2 SeV+\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eForward\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e23,177,522\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e23,161,169\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.93\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eReverse\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e23,177,522\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e23,161,169\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.93\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eOvBRCA2 SeV-\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eForward\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20,492,659\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e20,476,948\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.92\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eReverse\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e101\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e20,492,659\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e20,476,948\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e99.92\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eHuman transcriptome sequencing analysis\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"2\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c2\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003eGenes with more than 2-fold difference in expression\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eOvn\\u0026thinsp;\\u0026lt;\\u0026thinsp;Ov BRCA1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eOvn\\u0026thinsp;\\u0026lt;\\u0026thinsp;Ov BRCA2\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eGREM1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePAMR1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eC3\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCXCL1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCFB\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eTNC\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCXCL1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePLAU\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCXCL2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eGREM1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCSF3\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCXCL2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSEMA5A\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eC3\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eVCAM1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSEMA5A\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePTGS2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePAPPA\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePAPPA\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eHMCN1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIL6\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eRAB27B\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eRAB27B\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCFB\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCCL5\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCPE\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eHMCN1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCHL1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eLAMA5\\u003c/em\\u003e, \\u003cem\\u003eMIR4758\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eAADAC\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eFBLN2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eFBLN2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCCL2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eGBP4\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eC1R\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIGFBP6\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIGFBP6\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSPOCK1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eGBP4\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIL6\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePCOLCE\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCCL2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eTNC\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eNFKBIZ\\u003c/em\\u003e, \\u003cem\\u003eNXPE3\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eIGFBP1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eMMP1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eMMP1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSOX4\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCPE\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eANGPTL2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eTIMP3\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePRRX1\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eANGPTL2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eMMP16\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eCPZ\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eTMEM132A\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003ePDK4\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eOASL\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u003cem\\u003eSLC7A2\\u003c/em\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"2\\\"\\u003e\\u003cem\\u003eMMP1\\u003c/em\\u003e, \\u003cem\\u003eCCL2\\u003c/em\\u003e, \\u003cem\\u003eIGFBP6\\u003c/em\\u003e, \\u003cem\\u003eGREM1\\u003c/em\\u003e, \\u003cem\\u003eCFB\\u003c/em\\u003e, \\u003cem\\u003eCXCL1\\u003c/em\\u003e, \\u003cem\\u003eCXCL2\\u003c/em\\u003e, \\u003cem\\u003ePAPPA\\u003c/em\\u003e, \\u003cem\\u003eRAB27B\\u003c/em\\u003e, \\u003cem\\u003eFBLN2\\u003c/em\\u003e, and \\u003cem\\u003eC3\\u003c/em\\u003e were highly expressed in Ov BRCA1 or and Ov BRCA2 compared with their levels in Ovn cells\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eReal-time PCR\\u003c/em\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eMMP1\\u003c/em\\u003e, \\u003cem\\u003eRAB27B\\u003c/em\\u003e, CCL2, \\u003cem\\u003ePAPPA\\u003c/em\\u003e, \\u003cem\\u003eGREM1\\u003c/em\\u003e were highly expressed in Ov BRCA1 compared with their expression in Ovn, whereas \\u003cem\\u003eMMP1\\u003c/em\\u003e, \\u003cem\\u003eCXCL1, RAB27B, CCL2\\u003c/em\\u003e, \\u003cem\\u003ePAPPA\\u003c/em\\u003e and \\u003cem\\u003eGREM1\\u003c/em\\u003e were significantly highly expressed in Ov BRCA2 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eTo the best of our knowledge, this is the first study to successfully establish immortalized normal ovarian epithelial cells and ovarian epithelial cells from patients with germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e (g\\u003cem\\u003eBRCA1\\u003c/em\\u003e) or germline \\u003cem\\u003eBRCA2\\u003c/em\\u003e (g\\u003cem\\u003eBRCA2\\u003c/em\\u003e) mutations using SeV vectors carrying three immortalization genes. In chromosomal analyses, the presence of g\\u003cem\\u003eBRCA1\\u003c/em\\u003e or g\\u003cem\\u003eBRCA2\\u003c/em\\u003e mutations suggests that oncogenesis may be in progress because of SeV immortalization. Hence, these cells are useful for analyzing the oncogenic processes in ovarian cancer. Additionally, in the next-generation sequencing analysis, the cell lines with g\\u003cem\\u003eBRCA1\\u003c/em\\u003e or g\\u003cem\\u003eBRCA2\\u003c/em\\u003e mutation showed significantly high expression of genes such as \\u003cem\\u003eMMP1\\u003c/em\\u003e compared to the normal ovarian epithelium. This finding indicates that these genes may be important in oncogenesis.\\u003c/p\\u003e \\u003cp\\u003eChromosomal structural abnormalities were observed during the immortalization of ovarian epithelial cells using SeV. Their frequency was higher in Ov BRCA1 and Ov BRCA2 than in Ovn. Possible reasons for this include the instability of germline \\u003cem\\u003eBRCA\\u003c/em\\u003e mutations and a combination of the effects of SeV infection and passaging. Age-related factors may also be involved, as the age of the germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e carrier was 57 years and that of the germline \\u003cem\\u003eBRCA2\\u003c/em\\u003e carrier was 53 years. Nonetheless, as age increases, the age of oocytes and frequency of chromosomal abnormalities increase [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e]. Furthermore, ovarian cancer in g\\u003cem\\u003eBRCA1\\u003c/em\\u003e mutation carriers has been reported to occur more frequently at a younger age than in g\\u003cem\\u003eBRCA2\\u003c/em\\u003e mutation carriers [\\u003cspan additionalcitationids=\\\"CR14 CR15 CR16\\\" citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e]. However, as the present study was conducted on a single case, we will discuss the results after accumulating more cases and conducting similar studies in the future. We also examined vascular endothelial cells and stem cells and found no chromosomal abnormalities in these cells, suggesting that the changes may be unique to the ovaries (Oshimura, et al., Submitted). Konishi et al. [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e] established immortalized cells by transfecting human mammary epithelial cells with \\u003cem\\u003eh-TERT\\u003c/em\\u003e and generated \\u003cem\\u003eBRCA1\\u003c/em\\u003e-deficient cells to elucidate the development mechanism of breast cancer. According to their study results, haploinsufficiency of \\u003cem\\u003eBRCA1\\u003c/em\\u003e may accelerate the development of hereditary breast cancer by promoting genetic mutations. In that study, instability was induced by the deletion of \\u003cem\\u003eBRCA\\u003c/em\\u003e in normal cells. However, in the present study, we established immortalized normal ovarian epithelial cells and cells from ovarian epithelial specimens obtained from patients with germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e or germline \\u003cem\\u003eBRCA2\\u003c/em\\u003e and analyzed their properties. The use of these cell lines may contribute to the elucidation of oncogenic processes, as viral infections of unstable structures are thought to induce further chromosomal aberrations.\\u003c/p\\u003e \\u003cp\\u003eThe transcriptome analysis of each cell line and real-time PCR showed that \\u003cem\\u003eMMP1\\u003c/em\\u003e and several other genes were expressed at significantly high levels in the ovarian surface epithelial cells of \\u003cem\\u003eBRCA1\\u003c/em\\u003e and \\u003cem\\u003eBRCA2\\u003c/em\\u003e carriers. Germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e or \\u003cem\\u003eBRCA2\\u003c/em\\u003e carriers are at a higher risk of developing ovarian and breast cancers than non-carriers. MMPs are a type of extracellular matrix protein, and Yokoi et al. reported that \\u003cem\\u003eMMP1\\u003c/em\\u003e is an indicator of poor prognosis of ovarian cancer. Additionally, extracellular vesicles carrying \\u003cem\\u003eMMP1\\u003c/em\\u003e mRNA, which induce the apoptosis of mesothelial cells, were found in the ascites fluid of patients with cancer [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e]. In contrast, a meta-analysis of MMPs as risk factors for ovarian cancer reported no significant association; nonetheless, additional research is warranted [\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e]. In the future, we plan to confirm \\u003cem\\u003eMMP1\\u003c/em\\u003e expression using frozen tissues and ovarian serous carcinoma cell lines from g\\u003cem\\u003eBRCA1\\u003c/em\\u003e or g\\u003cem\\u003eBRCA2\\u003c/em\\u003e mutation carriers. In conclusion, the immortalized cell lines established in this study have the potential to elucidate a new oncogenic process by confirming the gene expression in normal and abnormal clones of the same cell line and via epigenomic analysis.\\u003c/p\\u003e \\u003cp\\u003eThis study has several strengths. First, SeV functions as an RNA in the cytoplasm and is not incorporated into the chromosomes of target cells, thus eliminating genotoxicity. Second, efficient gene transfer can be achieved in a short time with low infection titers or target cells. Third, transgenic cells do not produce infectious viral particles; hence, they can be safely used. Owing to these characteristics, SeV vectors are being developed as novel gene delivery systems, called cytoplasmic RNA vectors, for gene therapy and gene vaccines. They are being increasingly used as research tools in biotechnology and for developing bioproducts. In this study, we established immortalized cells using this method.\\u003c/p\\u003e \\u003cp\\u003eThis study also has some limitations. First, this study was based on the analysis of one case each, and it was not possible to fully examine whether the chromosomal abnormalities were unique to the case or ovarian cancer. However, the absence of abnormalities in other immortalized cell lines, such as vascular endothelial cells, suggests that these changes may be unique to ovarian cells. SeV vectors exist in the cytoplasm in the form of RNA throughout their life cycle and are not susceptible to insertional mutations or structural changes in chromosomes because of incorporation into the chromosomal genome. This is in contrast to retroviral vectors, which express genes by incorporation into chromosomes, or adenovirus vectors, adeno-associated virus vectors, and plasmid vectors, which exist as DNA in the cell nucleus away from chromosomes and can be incorporated into chromosomes at a certain frequency. Second, the SeV vector is temperature-sensitive, and temperature changes can introduce mutations at the amino acid level to facilitate the removal of the vector. Hence, the CO\\u003csub\\u003e2\\u003c/sub\\u003e incubator temperature must be set at 35\\u0026deg;C. Incubating cells at 37\\u0026deg;C may lead to changes in cell characteristics and senescence. Therefore, it is essential to handle these cells with utmost care and culture them in a CO\\u003csub\\u003e2\\u003c/sub\\u003e incubator with temperature control. Moreover, immunocytochemical staining yielded positive results for keratin in Ovn Passage\\u0026thinsp;+\\u0026thinsp;8, Ov BRCA1 Passage\\u0026thinsp;+\\u0026thinsp;9, and Ov BRCA2 Passage\\u0026thinsp;+\\u0026thinsp;9 cells and for vimentin in some cells (Online Resource 5). One possibility is that epithelial\\u0026ndash;mesenchymal transition occurred with cell passage. Another explanation is that some stromal cells may have strayed into the tumor cells. However, when we collect cells from patient specimens, we appropriately sample from the ovarian surface layer and usually confirm that they are epithelial cells.\\u003c/p\\u003e \\u003cp\\u003eIn this study, normal ovarian cells were successfully established using SeV, and ovarian epithelial cells were obtained from patients harboring germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e or \\u003cem\\u003eBRCA2\\u003c/em\\u003e mutations. These cell lines can be used in research on ovarian tumors in the future, to understand the mechanism of ovarian cancer development, as well as in research to identify drugs and new diagnostic markers. Additionally, our results suggest that \\u003cem\\u003eMMP1\\u003c/em\\u003e might be important during the development of ovarian cancer with \\u003cem\\u003eBRCA\\u003c/em\\u003e mutations.\\u003c/p\\u003e \\u003cp\\u003eAlthough the establishment of immortalized cells of the normal ovarian epithelium has been previously reported, we maintained ovarian epithelial cells generated from patients with g\\u003cem\\u003eBRCA1\\u003c/em\\u003e and g\\u003cem\\u003eBRCA2\\u003c/em\\u003e mutations in addition to normal ovarian epithelial cells. Furthermore, our original immortalization technology is safe and easy to use, which is a major advantage. Although some issues remain, this immortalization technology has the potential for further development and it is expected that its application will extend beyond cell lines to include other than ovarian cells. We plan to construct a library of immortalized cell lines in the future and establish a sustainable research infrastructure.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis work was supported by a research grant from the Tottori University Hospital and a Grant-in-Aid for Scientific Research (Grant number 21K16770) in 2021. \\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCompliance with Ethical Standards\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eDisclosure of potential conflicts of interest\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors, except Mitsuo Oshimura, declare that they have no financial or non-financial interests to disclose. \\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eResearch involving Human Participants\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthics Statement\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe study was approved by the Ethics Committee of Tottori University (IRB number: 20A179).\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eInformed consent\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eConsent to participate\\u003c/p\\u003e\\n\\u003cp\\u003eInformed consent was obtained from all individual participants included in the study.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent to publish\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor Contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eHiroaki Komatsu,\\u0026nbsp;Yasuhiro Kazuki, and\\u0026nbsp;Masayo Okawa\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003econceived the study.\\u0026nbsp;Hiroaki Komatsu\\u0026nbsp;and\\u0026nbsp;Masayo Okawa\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003edeveloped the statistical analysis plan and conducted statistical analyses. All authors contributed to the interpretation of the results.\\u0026nbsp;Hiroaki Komatsu\\u0026nbsp;drafted the original manuscript.\\u0026nbsp;Yasuhiro Kazuki\\u0026nbsp;and\\u0026nbsp;Masayo Okawa\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003esupervised the conduct of this study. All authors reviewed the manuscript draft and revised it critically for intellectual content. All authors approved the final version of the manuscript to be published.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData Availability\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors confirm that the data supporting the findings of this study are available within the article or its supplementary materials.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgments\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors are grateful to Mitsuhiko Ozaki, Ph.D., from the Department of Experimental Pathology, Department of Life Sciences, Faculty of Medicine, Tottori University, for providing guidance on the research methods.\\u003c/p\\u003e\\n\\u003cp\\u003eThis work was supported by a research grant from the Tottori University Hospital and a Grant-in-Aid for Scientific Research (21K16770) in 2021.\\u0026nbsp;\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eKurman RJ, Shih LM. The origin and pathogenesis of epithelial ovarian cancer: a proposed unifying theory. Am J Surg Pathol. 2010;34:433\\u0026ndash;43.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMogos RA, Popovici R, Tanase AE, et al. New approaches in ovarian cancer based on genetics and carcinogenesis hypotheses (Review). Exp Ther Med. 2022;23:423.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCancer Genome Atlas Research Network. Integrated genomic analyses of ovarian carcinoma. Nature. 2011;474:609\\u0026ndash;15.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature. 2012;490:61\\u0026ndash;70.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKonishi H, Mohseni M, Tamaki A, et al. Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells. Proc Natl Acad Sci U S A. 2011;108:17773\\u0026ndash;8.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eOhira M, Kikuchi E, Mizuta S, et al. Production of therapeutic iduronate-2-sulfatase enzyme with a novel single-stranded RNA virus vector. Genes Cells. 2021;26:891\\u0026ndash;904.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003ePotirat P, Wattanapanitch M, Kheolamai P, Issaragrisil S. Establishment of a human iPSC line (MUSIi007-A) from peripheral blood of normal individual using Sendai viral vectors. Stem Cell Res. 2018;32:43\\u0026ndash;6.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eTran TT, Nguyen TH, Nguyen TT, Nguyen XH. Establishment of a Vietnamese ethnicity induced pluripotent stem cell line (VRISGi001-A) from umbilical cord blood hematopoietic stem cells under a feeder-free system. Stem Cell Res. 2021;53:102345.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhou Z, Yoshimatsu S, Qian E, et al. Generation of a control human induced pluripotent stem cell line using the defective and persistent Sendai virus vector system. Stem Cell Res. 2021;56:102549.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLotila J, Hyv\\u0026auml;rinen T, Skottman H, Airas L, Narkilahti S, Hagman S. Establishment of a human induced pluripotent stem cell line (TAUi008-A) derived from a multiple sclerosis patient. Stem Cell Res. 2022;63:102865.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eItamochi H, Kato M, Nishimura M, et al. Establishment and characterization of a novel ovarian clear cell adenocarcinoma cell line, TU-OC-1, with a mutation in the PIK3CA gene. Hum Cell. 2013;26:121\\u0026ndash;7.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMishina T, Tabata N, Hayashi T, et al. Single-oocyte transcriptome analysis reveals aging-associated effects influenced by life stage and calorie restriction. Aging Cell. 2021;20:e13428.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eFord D, Easton DF. The genetics of breast and ovarian cancer. Br J Cancer. 1995;72:805\\u0026ndash;12.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eFord D, Easton DF, Stratton M, et al. Genetic heterogeneity and penetrance analysis of the BRCA1 and BRCA2 genes in breast cancer families. Am J Hum Genet. 1998;62:676\\u0026ndash;89.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSatagopan JM, Boyd J, Kauff ND, et al. Ovarian cancer risk in Ashkenazi Jewish carriers of BRCA1 and BRCA2 mutations. Clin Cancer Res. 2002;8:3776\\u0026ndash;81.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChen S, Iversen ES, Friebel T, et al. Characterization of BRCA1 and BRCA2 mutations in a large United States sample. J Clin Oncol. 2006;24:863\\u0026ndash;71.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMavaddat N, Peock S, Frost D, et al. Cancer risks for BRCA1 and BRCA2 mutation carriers: results from prospective analysis of EMBRACE. J Natl Cancer Inst. 2013;105:812\\u0026ndash;22.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eYokoi A, Yoshioka Y, Yamamoto Y, et al. Malignant extracellular vesicles carrying MMP1 mRNA facilitate peritoneal dissemination in ovarian cancer. Nat Commun. 2017;8:14470.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhu XM, Sun WF. Association between matrix metalloproteinases polymorphisms and ovarian cancer risk: A meta-analysis and systematic review. PLoS ONE. 2017;12:e0185456.\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\u003e\"},{\"header\":\"Tables\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eTable 1.\\u003c/strong\\u003e Chromosomal aberrations in the metaphase of each cell\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"2\\\"\\u003e\\n \\u003cp\\u003eCell line\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd rowspan=\\\"2\\\"\\u003e\\n \\u003cp\\u003eSeV\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd rowspan=\\\"2\\\"\\u003e\\n \\u003cp\\u003ePassage\\u003c/p\\u003e\\n \\u003cp\\u003eNo.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"5\\\"\\u003e\\n \\u003cp\\u003eChromosome No.\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"5\\\"\\u003e\\n \\u003cp\\u003eAberration\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd colspan=\\\"3\\\"\\u003e\\n \\u003cp\\u003eChromosome rearrangement\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e44\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e45\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e46\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e47\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eTotal\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003ectg/\\u003c/p\\u003e\\n \\u003cp\\u003eictg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003ectb/\\u003c/p\\u003e\\n \\u003cp\\u003eictb\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eexh\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eTotal\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eRatio\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eTranslocation, deletion, dicentric, etc.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eTotal\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eRatio\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"3\\\"\\u003e\\n \\u003cp\\u003eOvn normal\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eー\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eP+1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e24\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\u003cbr\\u003e\\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e+\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eP+5\\u003cbr\\u003e\\u0026nbsp;(early)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e46,XX,del(5q)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e4.0%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e+\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eP+25\\u003c/p\\u003e\\n \\u003cp\\u003e(late)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e24\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e46,XX,t(3;16)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,-22,-22,+2mar\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e2\\u003c/p\\u003e\\n \\u003cp\\u003e3/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e12%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd rowspan=\\\"2\\\"\\u003e\\n \\u003cp\\u003eOvn BRCA1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eー\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eP+3\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e23\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e4%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e47,XX,+mar\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e4%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e+\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eP+6\\u003cbr\\u003e\\u0026nbsp;(early)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e24\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e0%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e46,XX,add(11p),-19,+mar\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,t(6;11)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,del(7q)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,-7,-8,+2mar\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,add(5p)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,del(2p),del(3q)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,add(1q)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,del(3p),del(5q),del(7p)\\u003c/p\\u003e\\n \\u003cp\\u003e46,XX,del(Xq),add(11q)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e1\\u003c/p\\u003e\\n \\u003cp\\u003e9/25\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e36%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eTable 2.\\u0026nbsp;Read lengths and number of assembled transcripts for each cell line in human transcriptome sequencing analysis\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"595\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"20%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eSample\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.596638655462185%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eDirection\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.243697478991596%\\\" colspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOriginal\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.630252100840336%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eModified\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"23.529411764705884%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eRatio (%) (modified/original)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"35.58052434456929%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eRead length (bp)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.588014981273407%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eNo. of reads\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"34.831460674157306%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eNo. of reads\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"20%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvn SeV+\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.596638655462185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eForward\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.966386554621849%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"13.277310924369749%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,265,585\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.630252100840336%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,249,233\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"23.529411764705884%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.93\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"14.495798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eReverse\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.95798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"16.596638655462186%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,265,585\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.537815126050422%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,249,233\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.41176470588235%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.93\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"20%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvn SeV-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.596638655462185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eForward\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.966386554621849%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"13.277310924369749%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,629,854\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.630252100840336%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,611,040\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"23.529411764705884%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.92\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"14.495798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eReverse\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.95798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"16.596638655462186%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,629,854\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.537815126050422%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e22,611,040\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.41176470588235%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.92\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"20%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvBRCA1 SeV+\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.596638655462185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eForward\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.966386554621849%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"13.277310924369749%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,333,820\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.630252100840336%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,319,444\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"23.529411764705884%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.94\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"14.495798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eReverse\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.95798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"16.596638655462186%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,333,820\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.537815126050422%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,319,444\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.41176470588235%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.94\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"20%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvBRCA2 SeV+\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.596638655462185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eForward\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.966386554621849%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"13.277310924369749%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,177,522\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.630252100840336%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,161,169\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"23.529411764705884%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.93\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"14.495798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eReverse\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.95798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"16.596638655462186%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,177,522\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.537815126050422%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e23,161,169\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.41176470588235%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.93\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"20%\\\" rowspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvBRCA2 SeV-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"11.596638655462185%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eForward\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.966386554621849%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"13.277310924369749%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20,492,659\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"15.630252100840336%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20,476,948\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"23.529411764705884%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.92\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"14.495798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eReverse\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.95798319327731%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e101\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"16.596638655462186%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20,492,659\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"19.537815126050422%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e20,476,948\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"29.41176470588235%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e99.92\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 3\\u0026nbsp;\\u003c/strong\\u003e\\u003cem\\u003eHuman transcriptome sequencing analysis\\u003c/em\\u003e\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"100%\\\" colspan=\\\"2\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eGenes with more than 2-fold difference in expression\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvn \\u0026lt; Ov BRCA1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003eOvn \\u0026lt; Ov BRCA2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eGREM1\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003ePAMR1\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eC3\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCXCL1\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCFB\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eTNC\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCXCL1\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003ePLAU\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCXCL2\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eGREM1\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCSF3\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eCXCL2\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eSEMA5A\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eC3\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eVCAM1\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd width=\\\"50%\\\" valign=\\\"top\\\"\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eSEMA5A\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n 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\\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003e\\u003cem\\u003eMMP1\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;CCL2\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;IGFBP6\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;GREM1\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;CFB\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;CXCL1\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;CXCL2\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;PAPPA\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;RAB27B\\u003c/em\\u003e,\\u003cem\\u003e\\u0026nbsp;FBLN2\\u003c/em\\u003e, and \\u003cem\\u003eC3\\u003c/em\\u003e were highly expressed in Ov BRCA1 or and Ov BRCA2 compared with their levels in Ovn cells\\u003c/p\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":true,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"human-cell\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"huce\",\"sideBox\":\"Learn more about [Human Cell](http://link.springer.com/journal/13577)\",\"snPcode\":\"13577\",\"submissionUrl\":\"https://www.editorialmanager.com/huce/default2.aspx\",\"title\":\"Human Cell\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"ovarian epithelial cancer, carcinogenesis, Sendai virus, BRCA1/2\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-4012283/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-4012283/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eWe aimed to elucidate the mechanism underlying carcinogenesis by comparing normal and \\u003cem\\u003eBRCA1/2-\\u003c/em\\u003emutated ovarian epithelial cells established via Sendai virus-based immortalization. Ovarian epithelial cells (normal epithelium: Ovn; with germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e mutation: OvBRCA1; with germline \\u003cem\\u003eBRCA2\\u003c/em\\u003e mutation: OvBRCA2) were infected with Sendai virus vectors carrying three immortalization genes (\\u003cem\\u003eBmi-1\\u003c/em\\u003e, \\u003cem\\u003ehTERT\\u003c/em\\u003e, and \\u003cem\\u003eSV40T\\u003c/em\\u003e). The immunoreactivity of anti-epithelial cellular adhesion molecule (EpCAM) antibodies in each cell line and cells after 25 passages was confirmed using flow cytometry. Chromosomes were identified and karyotyped to detect numerical and structural abnormalities. Total RNA extracted from the cells was subjected to human transcriptome sequencing. Highly expressed genes in each cell line were confirmed using real-time polymerase chain reaction. Immortalization techniques allowed 25 or more passages of Ovn, OvBRCA1, and OvBRCA2 cells. No anti-EpCAM antibody reactions were observed in primary cultures or after long-term passages of each cell line. Structural abnormalities in the chromosomes were observed in each cell line; however, the abnormal chromosomes were successfully separated from the normal structures via cloning. Only normal cells from each cell line were cloned. \\u003cem\\u003eMMP1\\u003c/em\\u003e, \\u003cem\\u003eCCL2\\u003c/em\\u003e, and \\u003cem\\u003ePAPPA\\u003c/em\\u003e were more predominantly expressed in OvBRCA1 and OvBRCA2 cells than in Ovn cells. Immortalized ovarian cells derived from patients with germline \\u003cem\\u003eBRCA1\\u003c/em\\u003e or \\u003cem\\u003eBRCA2\\u003c/em\\u003e mutations showed substantially higher \\u003cem\\u003eMMP1\\u003c/em\\u003e expression than normal ovarian cells. However, the findings need to be validated in the future.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Characterization of immortalized ovarian epithelial cells with BRCA1/2 mutation\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-03-11 19:12:28\",\"doi\":\"10.21203/rs.3.rs-4012283/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"reviewerAgreed\",\"content\":\"\",\"date\":\"2024-03-07T09:59:19+00:00\",\"index\":0,\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2024-03-07T09:58:03+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2024-03-04T10:17:44+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Human Cell\",\"date\":\"2024-03-01T22:14:54+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"human-cell\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"huce\",\"sideBox\":\"Learn more about [Human Cell](http://link.springer.com/journal/13577)\",\"snPcode\":\"13577\",\"submissionUrl\":\"https://www.editorialmanager.com/huce/default2.aspx\",\"title\":\"Human Cell\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"b81486dd-c507-4ce3-9297-d0cd447912b7\",\"owner\":[],\"postedDate\":\"March 11th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-04-14T14:49:07+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-4012283\",\"link\":\"https://doi.org/10.1007/s13577-024-01064-z\",\"journal\":{\"identity\":\"human-cell\",\"isVorOnly\":false,\"title\":\"Human Cell\"},\"publishedOn\":\"2024-04-14 14:49:07\",\"publishedOnDateReadable\":\"April 14th, 2024\"},\"versionCreatedAt\":\"2024-03-11 19:12:28\",\"video\":\"\",\"vorDoi\":\"10.1007/s13577-024-01064-z\",\"vorDoiUrl\":\"https://doi.org/10.1007/s13577-024-01064-z\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4012283\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4012283\",\"identity\":\"rs-4012283\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}