Association of SNHG17/CDKN2B-AS1 polymorphisms with salpingitis and fallopian tube carcinoma susceptibility in the Chinese Han population.

Primary fallopian tube carcinoma (PFTC) is a kind of malignant fallopian tube carcinoma with low incidence, accounting for only 0.18–1.6% of female malignant tumors in reproductive system ( 1 ), but it is highly malignant, prone to pelvic and peritoneal dissemination and metastasis. Its main pathological type is high-grade serous fallopian tube carcinoma (HGSC) ( 2 ). Due to the application of opportunistic resection of fallopian tube, some early malignant lesions of fallopian tube have been found, and more and more evidence supports the theory of fallopian tube origin of epithelial ovarian cancer (EOC) and peritoneal serous adenocarcinoma ( 3 ), suggesting that the incidence rate of fallopian tube cancer may be underestimated. Studies have shown that genetic factors are associated with 20% of ovarian cancer, fallopian tube cancer and peritoneal cancer ( 4 ). Harmful genetic mutations in breast cancer 1 (BRCA1) and breast cancer 2 (BRCA2) are high-risk factors for HGSC. The risk of women with BRCA1 mutations is about 20–50%, while the risk of women with BRCA2 mutations is about 10–20%. Generally, these cancers occur earlier than sporadic cancers ( 2 , 5 - 7 ). In addition to breast cancer susceptibility gene ( BRCA ) gene, some genes with low penetrance may also induce HGSC, such as DNA mismatch repair genes related to Lynch syndrome [MutL homolog 1 (MLH1), MutS homolog 2 (MSH2), MutS homolog 6 (MSH6), PMS1 homolog 2 (PMS2), etc.], and DNA homologous repair pathway related molecules [BRCA1 interacting DNA helicase 1 (BRIP1), partner and localizer of BRCA2 (PALB2), RAD51 paralog C (RAD51C), RAD51 paralog D (RAD51D), BRCA1 Associated RING domain 1 (BARD1), etc.] ( 8 - 11 ). Single nucleotide polymorphism (SNP) refers to the DNA sequence polymorphism caused by single nucleotide variation in the genome. It is a specific nucleotide site in the genome. These sites have two different bases, and the population frequency of fewer bases is ≥1% ( 12 ). Studies have shown that SNP is closely related to a large number of physiological processes, such as cell differentiation and development, autophagy and apoptosis, and the occurrence and development of malignant tumors. SNPs are often double allelic polymorphisms, and their genetic differences are widely used to explain disease susceptibility ( 13 ). Studies have reported that the incidence of cervical cancer, ovarian cancer, endometrial cancer and other gynecological tumors are associated with SNP polymorphism. At present, there are few studies on the genetic susceptibility of fallopian tube cancer, but there have been many reports on the genetic susceptibility of SNP in gynecological tumors such as breast cancer, cervical cancer, endometrial cancer and ovarian cancer ( 14 ). Among HGSC related genes, the SNP at rs1165555 in the promoter region of BRCA1 gene is associated with sporadic breast cancer ( 12 ). SNPs at rs7797466 of PMS2 gene, rs6151662 of MSH3 gene and rs3136245 of MSH6 gene are associated with invasive ovarian cancer ( 15 ). Eight SNP sites of the BRIP1 gene are associated with the pathogenesis of cervical cancer ( 16 ). These studies suggest that SNP may also affect the genetic susceptibility of fallopian tube cancer. In addition to gene mutation, environmental factors are also important factors involved in tumor formation. A large number of studies have confirmed that chronic inflammation is closely related to the occurrence and development of tumors ( 17 ). Chronic pelvic inflammation can increase the risk of ovarian cancer by three times. Meanwhile, malignant tumors may be self-regulated by driving the inflammatory process. It can be seen that there is a clear correlation between chronic inflammation and malignant tumors ( 18 , 19 ). Compared with fallopian tube cancer, salpingitis has received more attention, which is mainly related to female infertility. With the gradual increase of invasive operations such as induced abortion, the number of patients with salpingitis also shows an upward trend ( 20 ). At present, it has been found that chronic inflammatory lesions of the fallopian tube may cause hyperplasia, hypertrophy, pseudocarcinomatous changes and anaplasia of the fallopian tube, and eventually induce fallopian tube cancer ( 1 ). Non-coding RNA (ncRNA) is a kind of RNA transcribed by DNA but not translated into protein. A large number of studies have confirmed that this kind of RNA can regulate the occurrence and development of diseases and is closely related to the pathophysiological processes of various diseases ( 21 ). In view of the extensive role and regulatory function of ncRNA, more and more studies began to focus on the relationship between SNP site of ncRNA and disease susceptibility. For example, rs10757274 site of CDKN2B-AS1 gene was confirmed to be related to the sensitivity of coronary atherosclerotic heart disease ( 13 ). SNP site rs7133268 of long-chain noncoding RNATHRIL is associated with susceptibility to cervical cancer ( 22 ). Many studies have shown that long-chain noncoding RNA SNHG17 and CDKN2B and CDKN2A antisense cis and trans regulatory RNA 1 (CDKN2B-AS1) are involved in the occurrence and development of a variety of tumors ( 23 , 24 ). However, its relationship with HGSC has not been studied. For the selection of specific SNPs—rs1578462 (SNHG17), rs3217992 (CDKN2B-AS1), and rs4977756 (CDKN2B-AS1)—the following rationale guided our choice: first, based on whole-genome sequencing results, these SNPs were identified as having potential functional significance, with preliminary evidence suggesting they may regulate the expression or activity of SNHG17 and CDKN2B-AS1, thereby influencing downstream pathways involved in inflammation and carcinogenesis. Second, the loci were selected to balance coverage of classical pathways (e.g., cell cycle regulation and inflammatory responses) with emerging mechanisms (e.g., ncRNA-mediated gene regulation), ensuring a comprehensive exploration of their potential roles in salpingitis and HGSC pathogenesis. Collectively, these SNPs were chosen to capture both known and novel biological insights into the disease processes under investigation. We present this article in accordance with the STREGA reporting checklist (available at https://gpm.amegroups.com/article/view/10.21037/gpm-25-9/rc ).

The study was conducted in accordance with the Declaration of Helsinki and its subsequent amendments. The study was approved by the Ethics Committee of West China Second University Hospital (No. 2016020), and written informed consent forms were obtained from all patients. The corresponding method is carried out in accordance with the approved guidelines. In this study, tubal tissue samples from the normal control group, tubal inflammation group, and tubal cancer group were collected from hospitalized patients who visited West China Second University Hospital, Sichuan University between June 2015 and December 2019, totaling 595 cases. Estimation of sample size: the Quanto software 1.2.3 version is used to estimate the sample size. The ratio between the fallopian tube inflammation group, HGSC group, and normal control group was 1:1:1. The relative risk is set as 2.0, and the statistical efficiency is 80% [trait TypeG dichotomous; analysis: case-control; case: control ratio 1:1; model: dominant; AlleleFreq =0.10; type I error rate 0.05; statistical power: 0.80; odds ratio (OR): 2.0]. Under the dominant genetic model, the sample size required is 180 cases. In this study, the fallopian tube tissue samples of the normal control group, salpingitis group and HGSC group were collected from West China Second Hospital of Sichuan University from June 2015 to December 2019. All fallopian tube tissues were removed surgically, then they were taken, fixed, dehydrated and embedded in paraffin, and finally made into wax blocks for preservation. All the above samples are from unrelated Han individuals. The patient’s medical records have complete basic information, clinical information and pathological examination results. The inclusion criteria of normal control group: (I) patients with non-inflammatory benign gynecological diseases who underwent salpingectomy; this includes: patients who underwent total hysterectomy and tubal ligation due to uterine fibroids; patients who underwent total hysterectomy and tubal ligation due to adenomyosis; patients who underwent removal of the affected ovary and fallopian tube due to benign ovarian cysts. (II) The samples were identified as complete and normal fallopian tube lumen structure by histopathological examination. Inflammatory diseases and malignant tumors were excluded ( 25 ). The inclusion criteria of salpingitis group: (I) patients with benign gynecological diseases who underwent salpingectomy; (II) the samples were confirmed to have chronic inflammatory changes of fallopian tubes by histopathological examination. Malignant tumors were strictly excluded. The inclusion criteria of HGSC group: the HGSC was diagnosed by histopathological examination, and other malignant tumors, autoimmune diseases and other metastatic tumors that may affect the final results were excluded ( 26 ). The pathology of salpingitis primarily involves thickening of the fallopian tube mucosa, congestion, interstitial oedema, extensive infiltration of neutrophils, and patchy desquamation of the mucosal epithelium leading to mutual adhesion. The lumen contains pus-like secretions formed by desquamated necrotic epithelium and a large number of inflammatory cells. The muscularis and serosa layers also exhibit varying degrees of inflammatory reactions. Lymphatic vessels and capillaries are dilated, filled with polymorphonuclear leukocytes and thrombi, and small abscesses form within the muscularis. The pathology of advanced serous ovarian cancer is mainly based on cystic and solid ovarian/fallopian tube masses accompanied by peritoneal dissemination and ascites. Papillary and micropapillary arrangements are accompanied by fibrous vascular axes. Destructive stromal invasion is accompanied by necrosis and lymphovascular invasion. We collected fallopian tube samples and extracted total DNA. The DNA was extracted from all sample paraffin sections, according to the instructions of the paraffin embedded tissue DNA rapid extraction kit of Beijing Tiangen Biochemical Technology Co., Ltd. (Beijing, China). The purity and concentration of the sample DNA was defined through the nanodrop full-automatic ultra-micro spectrophotometer. The DNA samples used for subsequent experiments should ensure the OD260/OD280 of the DNA sample should be 1.8±0.1, and the concentration should be greater than 50 ng/µL. SNPs were genotyped by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) ( Tables S1,S2 ). The size of the product fragments was distinguished by non-deformable polyacrylamide gel vertical electrophoresis. The PCR amplification was referred to the instructions for PCR rapid amplification kit of Beijing Biotechnology Co., Ltd. (Beijing, China). Genotyping was verified by gene sequencing and polyacrylamide gel electrophoresis (PAGE). The restriction enzymes used and the product information are shown in Table S3 . In this study, SNP sites rs1578462 of SNHG17 gene and rs3217992 and rs4977756 of CDKN2B-AS1 gene were selected. The datasets analysed during the study are available in the dbSNP repository, rs1578462 ( https://ncbi.nlm.nih.gov/snp/rs157846 ), rs3217992 ( https://ncbi.nlm.nih.gov/snp/rs3217992 ), and rs4977756 ( https://ncbi.nlm.nih.gov/snp/rs4977756 ). According to the dbSNP database information, specific amplification primers were designed by primer5 software. In order to ensure that the amplified fragment has specific restriction sites at the polymorphic sites, mismatches were introduced into the primers of rs3217992 and rs4977756, respectively. The primer sequences were synthesized by Chengdu Qingke Biotechnology Co., Ltd. (Chengdu, China), the names, primer sequences and fragment sizes of the candidate SNPs are shown in Tables S4,S5 . SPSS 22.0 statistical software was used to analyze the data. Genotype frequency was calculated. SNP stats software was used to calculate the frequency distribution of SNP genotypes in each group, and to analyze the relationship between SNP genotypes and clinical features such as International Federation of Gynecology and Obstetrics (FIGO) stage and lymph node metastasis. OR and 95% confidence interval (CI) were used to express the relative risk. Perform Hardy-Weinberg (H-W) genetic equilibrium tests on the genotype distributions of each candidate locus; compare the genotype frequency distributions of each group of SNP to clarify the correlation between polymorphism at each locus and salpingitis and tubal cancer; analyze the correlation between the frequency distribution of candidate loci and the clinical characteristics of tubal cancer. The genotype distribution of the selected samples from the normal control group, salpingitis group and HGSC group in the subjects was tested by H-W equilibrium (HWE) software ( https://cran.r-project.org/web/packages/HardyWeinberg/index.html ) to see if it conforms to the H-W balance. Chi-squared test was used to analyze the difference of genotype and allele distribution frequency between normal control group, salpingitis group and HGSC group. In addition, SHEsis online tool software ( http://analysis.bio-x.cn/myAnalysis.php ) was used to perform Haplotype analysis. P value <0.05 indicates statistically significant difference.

In our study, 595 samples were recruited and divided into 3 groups according to the histopathological diagnosis, including 206 cases of normal control, 194 cases of salpingitis and 195 cases of HGSC. The average ages were 54.4±8.30, 40.1±7.6, 54.4±8.39 years, respectively. In the HGSC group, there were 20 patients (10.3%) in stage I, 59 patients (30.3%) in stage II, 108 patients (55.4%) in stage III, 3 patients (1.4%) in stage IV, and the other 5 patients (2.6%) were of unknown stage. Among them, 52 patients had lymph node metastasis (26.7%) ( Table 1 ). Data are presented as number, mean ± SD, or n (%). FIGO, International Federation of Gynecology and Obstetrics; HGSC, high-grade serous fallopian tube carcinoma; SD, standard deviation. According to the screening principles described in the method, we selected three new SNPs, namely SNP of SNHG17 gene (rs1578462T/C) and SNP of CDKN2B-AS1 gene (rs3217992G/A and rs4977756G/A) as candidate subjects. The amplified products were directly sequenced to determine the genotyping of the loci, and the genotyping results were satisfactory ( Figures 1-3 ). Qualitative analysis of rs1578462. (A-C) Sequencing verification results of rs1578462 locus, red arrows represent polymorphic loci: (A) TT, (B) CT, (C) CC; (D) PAGE electrophoretic typing result: M-marker, TT [1,2,7,8,9], CC [4,5], CT [3,6]. PAGE, polyacrylamide gel electrophoresis. Qualitative analysis of rs3217992. (A-C) Sequencing verification results of rs3217992 locus, red arrows represent polymorphic loci: (A) GG, (B) AG, (C) AA; (D) PAGE electrophoretic typing result: M-marker, AG [1,4,6,9,11,12], GG [2,3,5,7,10], AA [8]. PAGE, polyacrylamide gel electrophoresis. Qualitative analysis of rs4977756. (A-C) Sequencing verification results of rs4977756 locus, red arrows represent polymorphic loci: (A) AA, (B) AG, (C) GG; (D) PAGE electrophoretic typing result: M-marker, AG [1,2,4,6], GG [3,7,8,9], AA [5]. PAGE, polyacrylamide gel electrophoresis. The frequency distribution of SNPs rs1578462T/C, rs3217992G/A and rs4977756A/G in the Chinese Han population was tested, the results showed that these SNPs are in HWE ( Table S6 ), indicating that the selected samples were from the same Mendelian population and were representative of the population. Their allele frequencies and genotype frequencies are shown in Table 2 . CI, confidence interval; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio; SNP, single nucleotide polymorphism. The frequency of CT/CC genotype was 40.5% in HGSC group, 50.5% in salpingitis group, which was lower than that in normal control group by 52.9%. The differences were both statistically significant (CT/CC vs. TT: OR =0.61, 95% CI: 0.41–0.90, P=0.01; CT/CC vs. TT: OR =0.67, 95% CI: 0.45–0.99, P=0.04). The frequency of TT/CT genotype in HGSC group (93.3%) was higher than that in normal control group (86.4%). The difference was also statistically significant (TT/CT vs. CC: OR =0.45, 95% CI: 0.23–0.90, P=0.02). The above results suggest that the CC and CT/CC genotypes at rs1578462 of SNHG17 may reduce the risk of HGSC. There was no significant difference in the frequency distribution of rs3217992G/A and rs4977756A/G polymorphisms of CDKN2B-AS1 among normal control group, salpingitis group and HGSC group (P>0.05), indicating that there was no correlation between these two polymorphisms and salpingitis and HGSC. We analyzed the linkage disequilibrium and haplotype of rs3217992G/A and rs4977756A/G polymorphisms of CDKN2B-AS1 according the typing results of the three groups. The D’ values of rs3217992 and rs4977756 were normal control vs. salpingitis (D’=0.512, r 2 =0.0428), normal control vs. fallopian tube cancer (D’=0.558, r 2 =0.0513) and salpingitis vs. ovarian cancer (D’=0.527, r 2 =0.0553), respectively, indicating moderate linkage disequilibrium. Haplotype analysis result showed that there was no significant difference among haplotypes ( Table S7 ). Next, we performed stratified analyses of polymorphisms for associations with age ( Table 3 ), FIGO stage ( Table 4 ), and Lymph node metastasis ( Table 5 ). CI, confidence interval; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio; SNP, single nucleotide polymorphism. CI, confidence interval; FIGO, International Federation of Gynecology and Obstetrics; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio; SNP, single nucleotide polymorphism. CI, confidence interval; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio; SNP, single nucleotide polymorphism. At rs3217992G/A locus, the frequency of AG genotype in HGSC group in FIGO stage III–IV (55.9%) was significantly higher than that in FIGO stage I–II (40.0%) (AG vs. GG: OR =2.17, 95% CI: 1.15–4.06, P=0.02). The frequency of AA/AG genotype in FIGO stage III–IV (69.4%) was significantly higher than that in FIGO stage I–II (52.5%). The difference was statistically significant (AA/AG vs. GG: OR =2.05, 95% CI: 1.13–3.72, P=0.02). For other clinical features, the difference was not statistically significant. At rs4977756A/G locus, the frequency of AG genotype in the group with lymph node metastasis (48.1%) was significantly higher than that in the group without lymph node metastasis (28.0%) (AG vs. AA: OR =0.42, 95% CI: 0.21–0.83, P=0.01). The frequency of AG/AG genotype in the group with lymph node metastasis (53.8%) was significantly higher than that in the group without lymph node metastasis (36.4%). The difference was statistically significant (AG/AG vs. AA: OR =0.49, 95% CI: 0.26–0.93, P=0.03), indicating that HGSC patients with AA/AG genotype were unlikely to have lymph node metastasis. In other clinical features, the difference was not statistically significant. There was no significant difference in genotype distribution among clinical features of rs1578462T/C locus. Combined with the results of clinical feature correlation analysis, we performed a joint analysis on the combination of candidate SNP sites rs1578462 + rs3217992 ( Table 6 ), rs1578462 + rs4977756 ( Table 7 ) and rs3217992 + rs4977756 ( Table 8 ) to evaluate the correlation between SNPs joint analysis and salpingitis and HGSC. CI, confidence interval; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio. CI, confidence interval; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio. CI, confidence interval; HGSC, high-grade serous fallopian tube carcinoma; OR, odds ratio. The frequency of rs1578462TT + rs3217992GG in salpingitis group (16.5%), which was lower than that in normal control group (22.3%). The difference was statistically significant (TT + GG vs. TT + AG/AA: OR =0.55, 95% CI: 0.31–0.99, P=0.04), indicating that the combined genotype of rs1578462TT + rs3217992GG may reduce the risk of salpingitis. The frequency of rs1578462CT/CC + rs3217992GG in HGSC group (16.4%), which was significantly lower than normal control group (23.3%). The difference was statistically significant (CT/CC + GG vs. TT + AG/AA: OR =0.45, 95% CI: 0.25–0.79, P=0.01), indicating that the combined genotype rs1578462CT/CC + rs3217992GG may reduce the risk of HGSC; the frequency of rs1578462CT/CC + rs3217992AG/AA in HGSC group (24.1%) was lower than that in normal control group (29.6%). The difference was statistically significant (CT/CC + AG/AA vs. TT + AG/AA: OR =0.52, 95% CI: 0.31–0.87, P=0.01). The frequency of rs1578462CT/CC + rs4977756AA in HGSC group (22.1%) lower than normal control group (35.0%), and the difference was statistically significant (CT/CC + AA vs. TT + AA: OR =0.48, 95% CI: 0.29–0.80, P=0.01). The frequency of rs3217992AG/AA + rs4977756AG/AG in HGSC group (21.5%), which was higher than that in normal control group (14.6%). The difference was statistically significant (AG/AA + AG/AG vs. GG + AA: OR =1.98, 95% CI: 1.04–3.76, P=0.04), indicating that the combined genotype rs3217992AG/AA + rs4977756AG/AG may increase the risk of HGSC. There was no significant difference in the frequency of other combinations in each group.

SNP is a sequence polymorphism caused by the change of a single nucleotide base. This sequence polymorphism is distributed throughout the genome, but uneven, with high genetic stability. At present, there are many classical methods for SNP typing, which can be roughly classified into detection technologies based on gel electrophoresis, such as RFLP-PCR, single strand conformation polymorphism (SSCP), denaturing gradient gel electrophoresis (DGGE), etc. the other detection technologies are based on molecular hybridization and fluorescent markers, such as TaqMan, etc. as well as high-throughput technologies developed in recent years, including sequencing chip, flight mass spectrometer technology and denaturing high performance liquid chromatography ( 27 ). RFLP-PCR was used in this study. This method is easy to operate, with stable typing results, high sensitivity, good repeatability and specificity. However, the restriction enzyme recognition sequences in the sequence polymorphic loci are limited, and the restriction enzyme digestion conditions are typical. Therefore, it is usually used in SNP typing at the selected location. For the detectability of formalin fixed paraffin embedded (FFPE) tissue samples, Stalberg et al. ( 28 ) used the archived wax blocks of patients with serous ovarian cancer to conduct SNP chip analysis. They verified the previously found genomic characteristics of ovarian cancer and proved that it is feasible to analyze genomic biomarkers on FFPE tissues. This study explored the association between SNP polymorphisms and the pathogenesis of HGSC. Our results showed that, in terms of single SNP genotype frequency distribution, the positive association was mainly reflected in the rs1578462 locus of the SNHG17 gene, suggesting that polymorphism at this locus may be related to the pathogenesis of HGSC. Joint analysis further supported the association with susceptibility: the combined genotypes of rs1578462 and rs3217992 (CT/CC + GG and CT/CC + AG/AA) were significantly less frequent in the HGSC group than in the normal control group (compared with TT + AG/AA genotype); similarly, the combined genotype of rs3217992 and rs4977756 (AG/AA + AG/AG) was significantly more frequent in the HGSC group than in the normal control group (compared with GG + AA genotype). These findings indicate that the above polymorphic combinations may be involved in the pathogenesis of HGSC. In contrast, the polymorphisms of CDKN2B-AS1 gene (rs3217992 and rs4977756) showed no significant differences in frequency distribution among the normal group, salpingitis group, and HGSC group, suggesting that these polymorphisms may not be associated with the occurrence of salpingitis or HGSC. Additionally, the genotypes of the three SNPs showed no frequency difference between the salpingitis group and the HGSC group, indicating they may not be related to the evolution from salpingitis to carcinoma. We further analyzed the association between SNPs and tumor progression-related clinical features (age of onset, FIGO stage, lymph node metastasis). For the rs1578462 locus of SNHG17, no significant differences in genotype frequency were observed among HGSC subgroups stratified by these clinical features, indicating that this locus is not associated with the age of onset, FIGO stage, or lymph node metastasis of HGSC. For CDKN2B-AS1 gene polymorphisms, however, specific associations with progression were noted: ❖ At the rs3217992 locus, the frequency of AG genotype and AA/AG genotype was significantly higher in FIGO stage III–IV than in stage I–II; ❖ At the rs4977756 locus, the frequency of AG genotype and AA/AG genotype was significantly lower in the non-lymph node metastasis group than in the lymph node metastasis group. These results suggest that rs3217992 and rs4977756 polymorphisms may be related to the progression of HGSC, as reflected by advanced FIGO stage and lymph node metastasis. Strengths: the use of paraffin blocks from actual cases and controls provided a reliable source of DNA. The selection of specific polymorphisms was based on prior knowledge and research, increasing the biological plausibility of the findings. The detailed analysis of multiple aspects such as susceptibility, grade, and metastasis provided a comprehensive view of the relationship between the genes SNP polymorphisms and HGSC. Limitations: the sample size, although relatively large with 195 HGSC cases and 206 controls, may still not be sufficient to capture all possible genetic and phenotypic variations. The study was focused on specific polymorphisms of two genes only, and other genetic factors or gene-gene interactions might have been overlooked. Additionally, the research was restricted to a Chinese Han population, limiting the generalizability to other ethnic groups. At present, there are a large number of studies on the correlation between SNP polymorphisms and the occurrence and clinical characteristics of malignant tumors, including in the field of gynecological tumors. Considering that most genome-wide association studies on EOC are mainly concentrated in European populations, Lawrenson et al. ( 29 ) selected 3,238 East Asian subjects diagnosed with EOC for SNP genotyping. They found that despite sharing some risk loci with European populations, East Asian populations still have some unique risk loci. This suggests that different populations have different genetic susceptibility to diseases, which also proves the necessity of SNP correlation verification for specific populations. Manichaikul et al. ( 30 ) conducted a genome-wide association study on women with EOC of African descent. They also found that there were differences in EOC variants between women of European and African descent. At the same time, they identified four new SNP sites related to EOC and six new SNP sites related to high-grade serous ovarian cancer. Most of these sites belong to non-coding regions. Lukács et al. ( 14 ) analyzed the correlation between rs2910164 locus of miR-146a and rs11614913 locus of mir-196a-2 and high-grade serous ovarian cancer, but there was no significant difference between the control group and the case group. The author also mentioned that increasing the number of cases may provide more accurate evidence about the relationship between the two. SNHG17 gene is located on chromosome 20 (20q11.23). In 2017, Ma et al. ( 23 ) first mentioned SNHG17 gene when studying the prognostic factors of colorectal cancer. They found that this molecule can promote the proliferation of colorectal cancer cells by silencing p57, which is an adverse prognostic factor. At present, most of the reports on this gene are functional studies. What can be clearly defined is that SNHG17 is related to the occurrence and development of a variety of cancers. SNHG17 can affect the expression of transcription factor Sp1 by acting as a competitive endogenous RNA of mir-876, thus promoting the malignant proliferation of tongue squamous cell carcinoma ( 31 ). It can promote the proliferation, migration and invasion of breast cancer by targeting mir-124-3p ( 32 ). Transcription factor YY1 can induce the expression of SNHG17 and activate Wnt by targeting mir-506-3p/ctnnb1 axis/β-catenin signaling pathway affects the development of glioma ( 33 ). It is also involved in the occurrence and development of non-small cell lung cancer, gastric cancer, melanoma and colorectal cancer. In addition, it has also been reported that SNHG17 is involved in the process of angiogenesis after ischemia-reperfusion, and its expression imbalance is also related to the susceptibility of type 2 diabetes ( 34 , 35 ). CDKN2B-AS1 gene is located in the CDKN2B-CDKN2A gene cluster on chromosome 9 (9p21), and its corresponding transcript is a functional RNA molecule, which can interact with polycomb inhibitory complexes protein regulator of cytokinesis 1 (PRC-1) and protein regulator of cytokinesis 2 (PRC-2) to induce the apparent silencing of other genes in the gene cluster. CDKN2B-AS1 as Cerna competes to bind miR-411-3p to promote apoptosis, inhibit proliferation, invasion and migration of ovarian cancer cells through HIF-1a/VEGF/P38 pathway. At the same time, miR-143-3p/SMAD3 axis can be targeted to indicate the prognosis of ovarian cancer. In hepatocellular carcinoma, CDKN2B-AS1 targets let-7c-5p/NAP1L1 axis to promote cell proliferation and metastasis ( 36 , 37 ). Since the region of this gene is an important genetic susceptibility site for cardiovascular disease, most studies focus on the correlation between SNPs of this gene and disease susceptibility. Xu et al. ( 13 ) conducted a meta-analysis on the correlation between rs10757274 locus of CDKN2B-AS1 gene and coronary atherosclerotic cardiomyopathy. Eleven studies included 52,209 subjects. They found that rs10757274 polymorphism may be used as a genetic marker of coronary heart disease, especially in the West Asian population. Zhao et al. ( 38 ) evaluated the correlation between the polymorphism of rs1333049 locus of CDKN2B-AS1 gene and hemorrhagic stroke and brain tumor. The study included 142 cases of hemorrhagic stroke, 115 cases of brain tumor and 494 controls. The results showed that there were differences in the correlation between the genotype of rs1333049 and the levels of low-density lipoprotein, high-density lipoprotein and total cholesterol in patients with hemorrhagic stroke. Meta-analysis showed that this locus was related to the risk of cerebrovascular disease. In addition, multiple SNP polymorphisms of CDKN2B-AS1 gene were also associated with susceptibility to type 2 diabetes, intracranial aneurysm, glaucoma, myocardial infarction, lung cancer and glioma. Interestingly, studies on CDKN2B-AS1 gene are mostly related to genetic susceptibility, while studies on snhg17 gene are mostly functional exploration. According to the data of dbSNP database, SNHG17 gene contains 4,111 SNP loci, while CDKN2B-AS1 gene contains 32759 SNP loci. The difference in the number of loci may be the reason for the different types of studies on the two genes. From the point of view of SNP locus, as mentioned above, there is no functional report on rs1578462 locus of SNHG17 gene. Some studies have studied the correlation between rs3217992 locus of CDKN2B-AS1 gene and plasma C-reactive protein (CRP) level in patients with coronary heart disease. The authors found that the increase of CRP concentration was mainly attributed to severe periodontitis, but not to the polymorphism of this locus ( 39 ). The allele of rs3217992 is also associated with an increased risk of pancreatic cancer ( 40 ). It is significantly correlated with anti-cyclic citrulline polypeptide antibody in patients with rheumatoid arthritis ( 41 ). It is related to myocardial infarction in Chinese Han population ( 42 ). In addition, in terms of functional research, rs3217992 has been proved to affect the expression regulation of CDKN2B-AS1 by miR-138-3p and miR-323b-5p ( 43 ). As for the rs4977756 locus of CDKN2B-AS1 gene, there are also some reports about its association with genetic susceptibility to disease. The correlation between the polymorphism of this locus and the risk of glioma is relatively clear, which has been proved by a large amount of evidence ( 44 , 45 ). Recent reports on the correlation between this site and the risk of glioma in Portugal and South India provide new support for this conclusion ( 46 , 47 ). Although some molecular evidences suggest the possible pathogenesis of oviduct cancer, the exact etiology of this tumor is not clear. It is certain that various genetic, endocrine and reproductive related factors are of great significance for the occurrence and development of oviduct cancer. Chronic inflammatory diseases have been considered as one of the causes of tissue malignant transformation, such as active hepatitis and liver cancer, ulcerative colitis and colon cancer, human papillomavirus infection and cervical cancer ( 48 ). Similarly, there are also studies that suggest that chronic pelvic inflammation is associated with the incidence of fallopian tube cancer. The early evidence is mainly the case reports of tuberculous salpingitis combined with primary salpingocarcinoma. Recent studies have found that salpingitis is closely related to the proliferation, hypertrophy, pseudocancerous changes and anaplasia of fallopian tubes ( 1 ). The significant association between the SNHG17 rs1578462 polymorphism and HGSC susceptibility suggests that this genetic variant may influence key molecular pathways involved in the initiation of the disease. The CC genotype potentially alters gene expression or protein function, leading to a decreased risk of developing HGSC. For CDKN2B-AS1 , the rs3217992 polymorphism’s relation to a higher grade of high-grade serous adenocarcinoma might imply its role in tumor progression and aggressiveness. The rs4977756 polymorphism’s link to lymph node metastasis indicates its potential impact on the cancer’s ability to spread. The combined effects of these polymorphisms on HGSC risk suggest complex interactions between different genetic variants in modulating the overall susceptibility and biological behavior of the cancer. Implications: these findings have significant implications for understanding the molecular basis of HGSC. They could potentially lead to the development of genetic screening tools for identifying individuals at a higher risk of developing HGSC. In the future, this could help in personalized prevention strategies and early detection. Moreover, the identified polymorphisms could serve as potential targets for the development of novel therapeutics. Actions needed: future research should aim to expand the sample size and include a more diverse population to validate and generalize the findings. Comprehensive genomic studies should be conducted to explore other genetic variants and their interactions with SNHG17 and CDKN2B-AS1 . Translational research is needed to translate these genetic findings into clinical applications, such as the development of reliable genetic tests and targeted therapies. Additionally, preclinical studies could be carried out to understand the exact molecular mechanisms by which these polymorphisms contribute to HGSC pathogenesis and progression.

Through analyzing DNA from 195 HGSC cases and 206 normal controls, we found the SNHG17 rs1578462 polymorphism was found to be significantly associated with HGSC susceptibility, with the CC genotype conferring a decreased risk. For CDKN2B-AS1 , the rs3217992 polymorphism was related to a higher grade of high-grade serous adenocarcinoma, and rs4977756 was associated with lymph node metastasis. Combinations of specific polymorphisms also correlated with HGSC risk. However, when examining the relationship between these gene polymorphisms and salpingitis, no statistically significant differences were observed. Although our study has several limitations, such as the sample size, though sizable, may not cover all variations, and the study was restricted to a Chinese Han population and focused on only two genes. These findings do have important implications. They could lead to the development of genetic screening tools for HGSC risk assessment and potentially inform personalized prevention and early detection strategies. Future research should expand the sample size and population diversity, conduct comprehensive genomic studies, and translate these genetic insights into clinical applications, including the development of genetic tests and targeted therapies, as well as preclinical investigations to elucidate the underlying molecular mechanisms.

The article’s supplementary files as