{"paper_id":"350b1b64-2a66-4d2f-aacd-98ddffa732f6","body_text":"LncRNA-DRNPC competitively binds miR-27a to inhibit EBV-LMP1-mediated ER (Endoplasmic reticulum) stress-metastasis in nasopharyngeal carcinoma (NPC) | 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 LncRNA-DRNPC competitively binds miR-27a to inhibit EBV-LMP1-mediated ER (Endoplasmic reticulum) stress-metastasis in nasopharyngeal carcinoma (NPC) Xianbao Cao, Lan Cai, Tianshu Li, Chun Feng, Jinqian Zhang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5330415/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background The details of the regulatory mechanism in metastasis of nasopharyngeal carcinoma related to lncRNAs are unclear. Methods The bio-functions of LncRNA-DRNPC-miR-27a-FAM172A interaction net in the relationships between ER stress, NPC invasion were investigated. Results There was high expression of miR-27a and low expression of LncRNA-DRNPC in NPC tumor tissues. LncRNA-DRNPC regulated miR-27a as a ceRNA, and miR-27a inhibited FAM172A expression, which participated in the regulation of ER stress-related NPC invasion induced by EBV-LMP1. Conclusion LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the NPC cell invasion associated with ER stress. DRNPC (Down-regulated in nasopharyngeal carcinoma) FAM172A gene ER (Endoplasmic reticulum) stress metastasis EBV (Epstein Barr virus) LMP1 (Latentmembraneprotein-1) Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Nasopharyngeal carcinoma (NPC) is an aggressive malignant tumor, especially for patients with recurrent or metastatic NPC. The median progression-free survival of NPC patients after treatment is only about seven months. However, during tumor metastasis progression, there is no standard method for second-line treatment, and the median overall survival in NPC is only about one year [ 1 , 2 ]. The etiology of NPC is complex, including genetic susceptibility, Epstein-Barrvirus (EBV) infection, and other environmental risk factors such as smoking, ingestion of preserved foods (Salted fish, etc.), and occupational exposure. The most important risk factor for NPC is EBV infection [ 3 ]. Recurrence and metastasis are the main reasons for the failure of NPC treatment. GP (Gemcitabine combined with cisplatin) chemotherapy is currently the first-line standard treatment for patients with recurrent or metastatic NPC, but its efficacy is limited and needs to be improved urgently [ 4 ]. The underlying reason for the poor prognosis of NPC patients is that the molecular mechanisms of invasion and metastasis are still unclear [ 5 ]. Therefore, exploring the molecular mechanism of NPC invasion and metastasis is always one of the key areas of tumor prevention and treatment, and clarifying the molecular mechanism of NPC metastasis will help to explore new molecular targets for metastatic NPC. Endoplasmic reticulum (ER) is the site of post-translational modification, folding, and assembly of membrane and secretory proteins in eukaryotic cells, and when these processes are affected, it can lead to Unfolded Protein Reaction (UPR) or misfolded proteins in ER. Reticulum accumulation causes ER stress (ERS, Endoplasmic reticulum stress), which is involved in the pathophysiological processes of various human tumors, including tumor migration, invasion and metastasis [ 6 ]. GRP78/Bip in NPC enables tumor cells to survive through various mechanisms, and is closely related to tumor drug resistance and proliferation, invasion and metastasis [ 7 ]. The interferon responses and ER stress could be induced by viral infection. Moreover, ER stress could by stimulated by viral polypeptides productions in the infection process of EBV [ 8 ]. The previous experimental data demonstrated the biological functions of FAM172A, which played a suppressor role in carcinoma, and was associated with ER stress [ 9 ]. In recently, LncRNA-DRNPC was found by our team, and it was confirmed that LncRNA-DRNPC could competitively combine with miR-27a. Thereby, the role of FAM172A in the relationships among ER stress, EBV infection, and metastasis progression was investigated. Meanwhile, the above biological function associated with the competitive combination among LncRNA-DRNPC, miR-27a, and FAM172A were explored in the metastasis progression of NPC related to ER stress induced by EBV infection. Materials and Methods Sample collection and detection of ncRNAs The tumor tissues of NPC patients (n=10) and their paired histologically normal tissue adjacent to the tumor (NAT) were collected from the First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology (Kunming, Yunnan Province, China). The Medical Research Ethics Committee of the First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology (Kunming 650032, Yunnan Province, China) approved this prospective study (No. KHYY-2021-18). The informed consent were provided to every participators, and who all signed a written informed consent form. Next-gene sequencing for noncoding RNAs of tissues Non-coding RNA-sequencing experiments were performed for extraction of RNAs from tumor tissues and their paired adjacent tissues. Then, the sequencing data were analyzed for confirming the differential miRNAs and lncRNAs with Ball gown software [10, 11]. Cell culture The poorly differentiated human NPC cell line CNE2 (CBP60003) were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). These cell lines were cultured in RPMI-1640 (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Thermo Fisher Scientific) at 37℃ in 5% CO 2 . qRT-PCR The expressions of miR-27a and lncRNA-DRNPC in tissues and cell lines were valuated utilizing assays of TaqMan miRNA, SYBR Green dye was used for the quantity of micorRNAs and lncRNAs with qRT-PCR method. The sequences of primers for miR-27a and lncRNA-DRNPC were shown in Supplementary Table 1 . The plasmid transfections The plasmid transfections were performed with jetPRIME agent (Polyplus-transfection, Illkirch-Graffenstaden, France) to achieve stable overexpression of LncRNA-DRNPC. The siRNA of LncRNA-DRNPC and mimics of miR-27a was purchased from Qiagen. The overexpression of LncRNA-DRNPC was constructed with pcDNA5/FRT/TO vector. RNA pull-down As the ceRNA was proved that miR-27a was regulated by lncRNA-DRNPC. Moreover, RNA pull-down was conducted to determine the combination between RISC complex and LncRNA-DRNPC. Subsequently, LncRNA-DRNPC was biotin-labeled after in vitro transcription with biotin RNA labeling mix (Roche). Furthermore, the above products were dealt with RNasE-Free DNase I (Roche), then incubated with the extraction of cell lines. After addition of streptavidin agarose beads (Invitrogen), western blot was performed to detect Ago 2 protein, and the expression level of miR-27a was tested with qRT-PCR from pellets of pulled down assay. Transwell assay For transwell invasion assay, cells were plated as described previously. Briefly, the wells were separated into upper and bottom chambers by a membrane with 8 µm round pores (Corning). The bottom of the upper chamber was coated with extracellular matrix gel mimicking the tumor micro-environment in vivo . Cells were plated on the top of the gel in the upper chamber while the media in the bottom chamber was supplemented with chemokines to induce cells penetrate the membrane and grow on the bottom surfaces of membrane. After 48 hrs of treatment, the number of cells growing in the bottom surface of membrane was quantified. Western blot analysis The protein electrophoreses were conducted on SDS-PAGE gel. Moreover, the blocked membrane transferred with proteins was dealt with skim milk and probed with primary antibody. The primary antibodies were utilized in our work, including β-actin (13E5) (Cell Signaling, Danvers, MA, USA; CST 4970S, dilution 1: 2,000), FAM172A (Abcam; ab 121364: dilution 1:2,000), anti-GRP78/BiP antibody (ab21685, dilution 1: 800; Abcam, Cambridge, MA, USA); anti-pATF2 (CST 40749, dilution 1: 800; Cell Signalling, Danvers, MA, USA); polyclonal rabbit anti-Human EMP-1 Antibody (aa35-63, LS-B12859, dilution 1: 600; Absolute Biotech, WA, USA). Finally, the chemiluminescence examination was performed based on the secondary HRP antibody using an exposed film. Statistics The data were shown as mean ± SD (Standard deviation). The differences between the two groups were analyzed with the Student’s t -test. Kruskal-Wallis ANOVA was used to analyze the abnormally distributed data among groups. SPSS software (version 18.0) was utilized to perform these statistical analyses. P < 0.05 was significant statistically. Results Differential miRNAs and lncRNAs expression analysis The differential expression of miRNAs was further valuated in NPC patients between the tumor tissues and their paired histologically normal tissues adjacent to the tumor (NAT) based on miRNA sequencing. Consequently, there were differentially expressed eleven miRNAs between the two groups, including nine upregulated expression of miRNAs and two downregulated miRNAs in FAM172A deficient mice ( Supplementary Table 2 ). Additionally, the relative miR-27a expression in tumor tissues of NPC patients was significantly enhanced than that in the paired normal tissues ( Figure 1A ). Moreover, after a series of screening works on lncRNAs sequencing, the differential lncRNAs expression analysis of the tumor tissues and their paired histologically normal tissues adjacent to the tumor (NAT) in NPC patients were performed, too. We found five lncRNAs, including three upregulated and two downregulated lncRNAs with stable expression differences ( Supplementary Table 3 ). After screening the lengths, adjacent coding genes, and other multiple conditions of the above ten lncRNAs, then these results show that NR_028080.1 had the full-length sequence in the open database. By querying the NCBI database, it was found that there was no report of NR_028080.1. Furthermore, the analysis of biological informatics confirmed that NR_028080.1 was the transcript coming from FAM172A gene, which was extensively studied by our team. Based on its expression level in tissues of NPC, we named NR_028080.1 as LncRNA-DRNPC (Down-regulated in nasopharyngeal carcinoma). Furthermore, qRT-PCR experiments were conducted to identify the differential expression of miRNAs and lncRNAs in tissues. These results demonstrated the high expression level of miR-27a ( Fig ure 1A ) and low expression level of LncRNA-DRNPC ( Fig ure 1B ) in tumor tissues coming from NPC patients than that in the control normal tissues. It was in line with the results of sequencing. LncRNA-DRNPC bound to miR-27a Whereafter, two biological informatics software DIANA-LncBase and miRcode, was used to predict the combination between the differential expressed miRNAs and lncRNAs selected from the above sequencing experiments. Then, the results indicated that miR-27a had binding sites of LncRNA-DRNPC ( Figure 2A ). Moreover, the overexpression ( Figure 2B ) and knockdown ( Figure 2C ) of LncRNA-DRNPC shifted miR-27a expression level. It suggested the targeted relationship between them, then the assay of RNA pull-down and dual luciferase reporter assay were conducted to confirm their interaction. According to the assay of dual luciferase reporter gene, the mimics of miR-27a and LncRNA-DRNPC-WT were co-transfected into NPC cell line CNE2. These results figured out the significantly decrease of the fluorescence induced by miR-27a ( Figure 2D ). Meanwhile, it identified LncRNA-DRNPC directly targeted miR-27a. Besides, Ago2 was detected in the mixed extraction of cell lines and LncRNA-DRNPC labeled with biotin according to RNA pull-down assay ( Figure 2E ), and it suggested the combination between RISC complex and LncRNA-DRNPC. In addition, the high expression level of miR-27a was found in DRNPC-pulled-down pellets based on qRT-PCR experiment ( Figure 2F ). These results suggested that as a ceRNA, LncRNA-DRNPC could regulate miR-27a. miR-27a inhibited FAM172A expression in NPC cell line The luciferase reporter gene system was further utilized to prove miR-27a straight target FAM172A in NPC cell line. Our results demonstrated that 3’ UTR luciferase activity of FAM172A downregulated clearly in CNE1 cell line after treatment with mimics of miR-27a ( Figure 3A ). Nevertheless, the mutation of FAM172A group did not exhibit significant alteration ( Figure 3B ). Meanwhile, miR-27a did not bind to the 3’ UTRs of GRP78/BiP , CHOP, ATF2, and EMP-1 (data not shown). Moreover, the role of miR-27a in FAM172A protein expression was explored. The results demonstrated that miR-27a suppressed FAM172A protein expression in CNE1 cell line ( Figure 3C ). In a word, it stated that miR-27a straight regulated the expression of FAM172A mRNA by binding to its 3′-UTR, and further affected its protein expression level. LncRNA-DRNPC competed with miR-27a for binding to FAM172A, and participated in the regulation of ER stress-related metastasis Through examining the expression levels of ER stress marker protein GRP78 and its key signal transduction pathway protein pATF2, we found that the LncRNA-DRNPC resulted in obviously increased expression levels of FAM172A, but the remarkably decreased of GRP78 and pATF2 expression level in CNE2 cell line. On the contrary, miR-27a caused significantly increased expression levels of GRP78 and pATF2 through suppressing FAM172A expression. However, the above biological function of miR-27a could be reversed by LncRNA-DRNPC ( Figure 4A ). Furthermore, the transwell invasion assay was conducted to explore the bio-function of above interaction net. It was declared that LncRNA-DRNPC effectively inhibited cell invasion of NPC cell line CNE2 ( P <0.01). On the contrary, miR-27a resulted in a significant promotion of cell invasion of NPC cell line CNE2 ( P <0.01). However, the cell invasion induced by miR-27a could be reversed by LncRNA-DRNPC ( Figure 4B and 4C ). These results indicated that LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the cell invasion associated with ER stress in NPC cell line CNE2. EBV-LMP1-mediated ER stress-metastasis associated with the interaction net of LncRNA-DRNPC-miR-27a-FAM172A Furthermore, qRT-PCR experiments were conducted to identify the differential expression of miRNAs and lncRNAs in CNE2. These results demonstrated the high expression level of miR-27a ( Fig ure 5A ) and low expression level of LncRNA-DRNPC ( Fig ure 5B ) in the group of treatment with EBV-LMP1. EBV-LMP1 (Latent membrane protein 1) is the major oncoprotein encoded by EBV, and contributes to the metastatic of NPC. Moreover，LMP1 protein induced down-expression of FAM172A, over-expression of the ERS marker protein GRP78, and the up-regulated of EMP-1. EMP-1 was the key regulator of tumor invasion ( Fig ure 6A ). Furthermore, the results of transwell invasion assay demonstrated that EBV-LMP1 promoted ERS in NPC cell line CNE2, and induced cell invasion of NPC. However, the cell invasion induced by EBV-LMP1 could be reversed by LncRNA-DRNPC ( Figure 6B and 6C ). These results indicated that LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the NPC cell invasion associated with ER stress. Discussion Although platinum-containing induction chemotherapy and radiotherapy is used as the first-line standard treatment, the median progression-free survival of NPC patients after treatment is only about seven months. During tumor metastasis progression, there is no standard method for second-line treatment, and the median overall survival in NPC is only about one year. Additionally, more than 130,000 new cases of NPC were diagnosed worldwide in 2020, nearly half of which occurred in China, especially in southern China [ 1 ]. The incidence and mortality of NPC in China are higher than the global average, and the death cases account for about 40% of all deaths from NPC in the world [ 2 ]. Therefore, to provide the new treatment strategies for patients with NPC is the crucial significance for improving the prognosis of advanced NPC. The principal oncoprotein of EBV (Epstein-Barr Virus), LMP1 (Latent Membrane Protein 1) is the superfamily receptor of TNF (Tumor Necrosis Factor) [ 12 , 13 ]. The features of LMP1 figure out its constitutive activity, and PRRs (Pathogen Recognition Receptors) functions, such as NRF2 of the master antioxidative transcription factor, IRFs subset. Moreover, its signal transduction lead to the activation of Akt, AP1, and NF-κB, and include the utilization of kinase cascades, adaptors, and TRAFs [ 12 – 22 ]. Furthermore, LMP1 is a pleiotropic functional factor involved in the balance and reprogram in the broad spectrum of cellular functions, including telomere elongation, immune defenses, extracellular vehicles, DNA damage response, epigenetics, metabolism, ubiquitin machinery [ 13 – 18 ]. Functionally, LMP1 promote the development of the tumor microenvironment, metastasis, invasion, cell survival and proliferation, anchorage-independent cell growth, angiogenesis, as well as oncogenic transformation [ 18 – 22 ]. Therefore, we explored the relationship between LMP1 and metastasis associated with ER stress. EMP1 was the key role in metastatic recurrence, then it was utilized in our work to confirm metastasis status [ 23 ]. At first, the LncRNAs closely related to nasopharyngeal carcinoma was screened by our research group through the next generation sequencing and proved to be the transcript of the tumor suppressor gene FAM172A in NPC, so it was named DRNPC. Consequently, the next-genesequencing experiments of noncoding RNAs were conducted based on the NPC tissues of patients. The differential microRNAs and lncRNA were found and identified, including miR-27a and LncRNA-DRNPC. In addition, our results further confirmed that miR-27a had binding sites with LncRNA-DRNPC, and LncRNA-DRNPC could regulate miR-27a as a ceRNA. At the mean time, miR-27a inhibited FAM172A expression in NPC cell line. Moreover, miR-27a resulted in a significant promotion of cell invasion of NPC cell line CNE2. However, the cell invasion induced by miR-27a could be reversed by LncRNA-DRNPC. LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the cell invasion associated with ER stress in NPC cell line CNE2. Secondly, we found that EBV-LMP1 promoted ERS in NPC cell line CNE2, and induced cell invasion of NPC. However, the cell invasion induced by EBV-LMP1 could be reversed by LncRNA-DRNPC. Finally, LncRNA-DRNPC inhibited EBV infection-associated nasopharyngeal carcinoma cell metastasis and endoplasmic reticulum stress, the metastasis and ER stress were positively correlated in NPC. LncRNA-DRNPC can bind to FAM172A, while miR-27a can degrade its target gene FAM172A. Therefore, we speculated that LncRNA-DRNPC might inhibit the progression of the expression of the target gene FAM172A through competitive binding with miR-27a, and reverse the progression of LMP1 encoded by Epstein Barr virus (EBV). The endoplasmic reticulum stress induced by LMP1 can promote the metastasis process of nasopharyngeal carcinoma cells. This project aims to further explore the molecular mechanism of LncRNA-DRNPC's regulation of EBV-LMP1-mediated ER stress-nasopharyngeal carcinoma cell invasion through the construction of over-expression of LncRNA-DRNPC and miR-27a silencing. To elucidate the mechanism of LncRNA-DRNPC as a ceRNA of miR-27a in the occurrence and development of EBV infection-associated nasopharyngeal carcinoma, and to provide key new molecular targets for the prevention and treatment of EBV infection-associated nasopharyngeal carcinoma. Conclusion Our work illustrated that LncRNA-DRNPC competitively binds miR-27a to inhibit EBV-LMP1-mediated ER (Endoplasmic reticulum) stress-metastasis in nasopharyngeal carcinoma (NPC). Declarations Ethics approval and consent to participate This study was approved by the Medical Research Ethics Committee of the The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology. Each patient was gave informed con[1]sent and signed a written informed consent form. Consent for publication Not applicable. Availability of data and materials The data used to support the findings of this study are included within the article. Competing interests The authors declare that they have no competing interests. Authors' contributions XBC, LC, and JQZ contributed to the conception of the study; XBC, and LC contributed significantly to performing these experiments; XBC, LC, THL, CF, and JQZ performed the data analyses and wrote the manuscript; XBC, CL, and JQZ helped perform the analysis with constructive discussions. Funding This work was supported by grants from the National Natural Science Foundation of China (No. 82360224),the Open Project of Yunnan Provincial Key Laboratory of Clinical Virology (202205AG070053), the Key Basic Research Program of Yunnan Province (No. 202201AS070065), and Kunming University of Science and Technology Medical Joint Project (KUST-KH2022038Y). The study sponsors had no involvement in the work. Acknowledgements Not applicable. 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Supplementary Files SupplementaryTables.doc Cite Share Download PDF Status: Posted Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-5330415\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":370629804,\"identity\":\"75a31d23-4b30-40bb-a700-7a14bb49528d\",\"order_by\":0,\"name\":\"Xianbao Cao\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Xianbao\",\"middleName\":\"\",\"lastName\":\"Cao\",\"suffix\":\"\"},{\"id\":370629805,\"identity\":\"5432cac0-d6e7-41b4-b553-618f1b244740\",\"order_by\":1,\"name\":\"Lan Cai\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"The Yongchuan District People's Hospital in Chongqing, Chongqing Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Lan\",\"middleName\":\"\",\"lastName\":\"Cai\",\"suffix\":\"\"},{\"id\":370629806,\"identity\":\"667dc3ce-6024-427c-9c6c-59852343490f\",\"order_by\":2,\"name\":\"Tianshu Li\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Tianshu\",\"middleName\":\"\",\"lastName\":\"Li\",\"suffix\":\"\"},{\"id\":370629807,\"identity\":\"a9f56833-4edc-4bd3-932d-4634a7bfc921\",\"order_by\":3,\"name\":\"Chun Feng\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Chun\",\"middleName\":\"\",\"lastName\":\"Feng\",\"suffix\":\"\"},{\"id\":370629808,\"identity\":\"cbbff10c-21de-44f3-a625-1b81403f62ef\",\"order_by\":4,\"name\":\"Jinqian Zhang\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA00lEQVRIiWNgGAWjYBACxmYowwDIfpBQUUOaFmaDB2eOkWAdUAub5MMWZsIqmdt5D7/mqbljt529x6wisYGNgb+9O4GAw/jSrHmOPUve2XPG7EbiDhkGiTNnNxDQwmNmnMN2ONngRg5Qyxk2BgOJXGK0/INoKUhsYyZKi/Hj3LbDdiAtDMRqMWP+23c4weDMsWKJhDPHeAj6xbD/jPHHGd8O2xscb9748UdFjRx/ey8BLQ0MbBJAOrEBKsCDVzkIyAOj5gOQtieochSMglEwCkYuAADV4ErddOEIxgAAAABJRU5ErkJggg==\",\"orcid\":\"\",\"institution\":\"The First People’s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Jinqian\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-10-25 07:38:17\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-5330415/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-5330415/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":68353316,\"identity\":\"607a96c4-140a-40eb-904f-37c7c13ebfd9\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:03:20\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":46873,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDifferential miRNAs and lncRNAs expression analysis.\\u003c/p\\u003e\\n\\u003cp\\u003eThe differential expression of miRNAs was further valuated in NPC patients between the tumor tissues and their paired histologically normal tissues adjacent to the tumor (NAT) based on miRNA sequencing. Additionally, the relative miR-27a expression in tumor tissues of NPC patients was significantly enhanced than that in the paired normal tissues (\\u003cstrong\\u003eA\\u003c/strong\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, qRT-PCR experiments were conducted to identify the differential expression of miRNAs and lncRNAs in tissues. These results demonstrated the high expression level of miR-27a \\u003cstrong\\u003eA\\u003c/strong\\u003e) and low expression level of LncRNA-DRNPC (\\u003cstrong\\u003eB\\u003c/strong\\u003e) in tumor tissues coming from NPC patients than that in the control normal tissues. It was in line with the results of sequencing.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/22f7107774c16f951bab8db7.png\"},{\"id\":68354838,\"identity\":\"49d28883-790e-4b79-9fa0-1448eb5d24a0\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:11:19\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":258848,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eLncRNA-DRNPC bound to miR-27a.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eOur results indicated that miR-27a had binding sites of LncRNA-DRNPC (\\u003cstrong\\u003eA\\u003c/strong\\u003e). Moreover, the overexpression (\\u003cstrong\\u003eB\\u003c/strong\\u003e) and knockdown (\\u003cstrong\\u003eC\\u003c/strong\\u003e) of LncRNA-DRNPC shifted miR-27a expression level. It suggested the targeted relationship between them, then the assay of RNA pull-down and dual luciferase reporter assay were conducted to confirm their interaction. According to the assay of dual luciferase reporter gene, the mimics of miR-27a and LncRNA-DRNPC-WT were co-transfected into NPC cell line CNE2. These results figured out the significantly decrease of the fluorescence induced by miR-27a (\\u003cstrong\\u003eD\\u003c/strong\\u003e). Meanwhile, it identified LncRNA-DRNPC directly targeted miR-27a. Besides, Ago2 was detected in the mixed extraction of cell lines and LncRNA-DRNPC labeled with biotin according to RNA pull-down assay (\\u003cstrong\\u003eE\\u003c/strong\\u003e), and it suggested the combination between RISC complex and LncRNA-DRNPC. In addition, the high expression level of miR-27a was found in DRNPC-pulled-down pellets based on qRT-PCR experiment (\\u003cstrong\\u003eF\\u003c/strong\\u003e).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/29834b25788348d92fb9e818.png\"},{\"id\":68353312,\"identity\":\"08c3ad8f-6784-4693-a1f2-309a59a38ab1\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:03:19\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":76751,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003emiR-27a inhibited FAM172A expression in NPC cell line.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe 3’ UTR luciferase activity of FAM172A downregulated clearly in CNE1 cell line after treatment with mimics of miR-27a (\\u003cstrong\\u003eA\\u003c/strong\\u003e). Nevertheless, the mutation of FAM172A group did not exhibit significant alteration (\\u003cstrong\\u003eB\\u003c/strong\\u003e). Moreover, miR-27a suppressed FAM172A protein expression in CNE1 cell line (\\u003cstrong\\u003eC\\u003c/strong\\u003e).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/c2c6737ac6cb2c723d29432d.png\"},{\"id\":68354839,\"identity\":\"bc8aeee8-1919-4dbb-9df8-7b0be5957c71\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:11:19\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1566008,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eLncRNA-DRNPC competed with miR-27a for binding to FAM172A, and participated in the regulation of ER stress-related metastasis.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThrough examining the expression levels of ER stress marker protein GRP78 and its key signal transduction pathway protein pATF2, we found that the LncRNA-DRNPC resulted in obviously increased expression levels of FAM172A, but the remarkably decreased of GRP78 and pATF2 expression level in CNE2 cell line. On the contrary, miR-27a caused significantly increased expression levels of GRP78 and pATF2 through suppressing FAM172A expression. However, the above biological function of miR-27a could be reversed by LncRNA-DRNPC (\\u003cstrong\\u003eA\\u003c/strong\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, LncRNA-DRNPC effectively inhibited cell invasion of NPC cell line CNE2. On the contrary, miR-27a resulted in a significant promotion of cell invasion of NPC cell line CNE2. However, the cell invasion induced by miR-27a could be reversed by LncRNA-DRNPC (\\u003cstrong\\u003eB and C\\u003c/strong\\u003e).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/c8d79a5b44aa402b7c6800aa.png\"},{\"id\":68353311,\"identity\":\"911c2c62-c23b-402f-adb1-956dbb4c0223\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:03:19\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":36803,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eEBV-LMP1-mediated ER stress-metastasis associated with the interaction net of LncRNA-DRNPC-miR-27a-FAM172A.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, qRT-PCR experiments were conducted to identify the differential expression of miRNAs and lncRNAs in CNE2. These results demonstrated the high expression level of miR-27a (\\u003cstrong\\u003eA\\u003c/strong\\u003e) and low expression level of LncRNA-DRNPC (\\u003cstrong\\u003eB\\u003c/strong\\u003e) in the group of treatment with EBV-LMP1. EBV-LMP1 (Latent membrane protein 1) is the major oncoprotein encoded by EBV, and contributes to the metastatic of NPC.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/ab33ec97ce4d67ff30f8590b.png\"},{\"id\":68353313,\"identity\":\"d2fee602-261e-41ac-b57e-4df89874d481\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:03:19\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1307983,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cstrong\\u003eEBV-LMP1-mediated ER stress-metastasis associated with the interaction net of LncRNA-DRNPC-miR-27a-FAM172A.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eMoreover，LMP1 protein induced down-expression of FAM172A, over-expression of the ERS marker protein GRP78, and the up-regulated of EMP-1. EMP-1 was the key regulator of tumor invasion (\\u003cstrong\\u003eFigure A\\u003c/strong\\u003e). Furthermore, the results of transwell invasion assay demonstrated that EBV-LMP1 promoted ERS in NPC cell line CNE2, and induced cell invasion of NPC. However, the cell invasion induced by EBV-LMP1 could be reversed by LncRNA-DRNPC (\\u003cstrong\\u003eB and C\\u003c/strong\\u003e).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/8f02e4875275785105ba9f4e.png\"},{\"id\":69163873,\"identity\":\"a36c1d06-2e9c-49d1-aaeb-a2cacec0406d\",\"added_by\":\"auto\",\"created_at\":\"2024-11-17 01:01:24\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":4189626,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/6e15f096-0388-40bd-a714-fa56c942b73b.pdf\"},{\"id\":68353317,\"identity\":\"384a257c-bd66-43ce-a746-001f0ed3f451\",\"added_by\":\"auto\",\"created_at\":\"2024-11-06 11:03:20\",\"extension\":\"doc\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":39424,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"SupplementaryTables.doc\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5330415/v1/c73511159362d24078971a72.doc\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"LncRNA-DRNPC competitively binds miR-27a to inhibit EBV-LMP1-mediated ER (Endoplasmic reticulum) stress-metastasis in nasopharyngeal carcinoma (NPC)\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eNasopharyngeal carcinoma (NPC) is an aggressive malignant tumor, especially for patients with recurrent or metastatic NPC. The median progression-free survival of NPC patients after treatment is only about seven months. However, during tumor metastasis progression, there is no standard method for second-line treatment, and the median overall survival in NPC is only about one year [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. The etiology of NPC is complex, including genetic susceptibility, Epstein-Barrvirus (EBV) infection, and other environmental risk factors such as smoking, ingestion of preserved foods (Salted fish, etc.), and occupational exposure. The most important risk factor for NPC is EBV infection [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eRecurrence and metastasis are the main reasons for the failure of NPC treatment. GP (Gemcitabine combined with cisplatin) chemotherapy is currently the first-line standard treatment for patients with recurrent or metastatic NPC, but its efficacy is limited and needs to be improved urgently [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. The underlying reason for the poor prognosis of NPC patients is that the molecular mechanisms of invasion and metastasis are still unclear [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. Therefore, exploring the molecular mechanism of NPC invasion and metastasis is always one of the key areas of tumor prevention and treatment, and clarifying the molecular mechanism of NPC metastasis will help to explore new molecular targets for metastatic NPC.\\u003c/p\\u003e \\u003cp\\u003eEndoplasmic reticulum (ER) is the site of post-translational modification, folding, and assembly of membrane and secretory proteins in eukaryotic cells, and when these processes are affected, it can lead to Unfolded Protein Reaction (UPR) or misfolded proteins in ER. Reticulum accumulation causes ER stress (ERS, Endoplasmic reticulum stress), which is involved in the pathophysiological processes of various human tumors, including tumor migration, invasion and metastasis [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e]. GRP78/Bip in NPC enables tumor cells to survive through various mechanisms, and is closely related to tumor drug resistance and proliferation, invasion and metastasis [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e]. The interferon responses and ER stress could be induced by viral infection. Moreover, ER stress could by stimulated by viral polypeptides productions in the infection process of EBV [\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe previous experimental data demonstrated the biological functions of FAM172A, which played a suppressor role in carcinoma, and was associated with ER stress [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. In recently, LncRNA-DRNPC was found by our team, and it was confirmed that LncRNA-DRNPC could competitively combine with miR-27a. Thereby, the role of FAM172A in the relationships among ER stress, EBV infection, and metastasis progression was investigated. Meanwhile, the above biological function associated with the competitive combination among LncRNA-DRNPC, miR-27a, and FAM172A were explored in the metastasis progression of NPC related to ER stress induced by EBV infection.\\u003c/p\\u003e\"},{\"header\":\"Materials and Methods\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eSample collection and detection of ncRNAs\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe tumor tissues of NPC patients (n=10) and their paired histologically normal tissue adjacent to the tumor (NAT) were collected from the First People\\u0026rsquo;s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology (Kunming, Yunnan Province, China).\\u003c/p\\u003e\\n\\u003cp\\u003eThe Medical Research Ethics Committee of the First People\\u0026rsquo;s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology (Kunming 650032, Yunnan Province, China) approved this prospective study (No. KHYY-2021-18). The informed consent were provided to every participators, and who all signed a written informed consent form. \\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eNext-gene sequencing for noncoding RNAs of tissues\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNon-coding RNA-sequencing experiments were performed for extraction of RNAs from tumor tissues and their paired adjacent tissues. Then, the sequencing data were analyzed for confirming the differential miRNAs and lncRNAs with Ball gown software [10, 11]. \\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eCell culture\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe poorly differentiated human NPC cell line CNE2 (CBP60003) were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). These cell lines were cultured in RPMI-1640 (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Thermo Fisher Scientific) at 37℃ in 5% CO\\u003csub\\u003e2\\u003c/sub\\u003e.\\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eqRT-PCR\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe expressions of miR-27a and lncRNA-DRNPC in tissues and cell lines were valuated utilizing assays of TaqMan miRNA, SYBR Green dye was used for the quantity of micorRNAs and lncRNAs with qRT-PCR method. The sequences of primers for miR-27a and lncRNA-DRNPC were shown in \\u003cstrong\\u003eSupplementary\\u003c/strong\\u003e \\u003cstrong\\u003eTable 1\\u003c/strong\\u003e.\\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eThe plasmid transfections \\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe plasmid transfections were performed with jetPRIME agent (Polyplus-transfection, Illkirch-Graffenstaden, France) to achieve stable overexpression of LncRNA-DRNPC. The siRNA of LncRNA-DRNPC and mimics of miR-27a was purchased from Qiagen. The overexpression of LncRNA-DRNPC was constructed with pcDNA5/FRT/TO vector.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003e \\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eRNA pull-down \\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAs the ceRNA was proved that miR-27a was regulated by lncRNA-DRNPC. Moreover, RNA pull-down was conducted to determine the combination between RISC complex and LncRNA-DRNPC. Subsequently, LncRNA-DRNPC was biotin-labeled after \\u003cem\\u003ein vitro\\u003c/em\\u003e transcription with biotin RNA labeling mix (Roche). Furthermore, the above products were dealt with RNasE-Free DNase I (Roche), then incubated with the extraction of cell lines. After addition of streptavidin agarose beads (Invitrogen), western blot was performed to detect Ago 2 protein, and the expression level of miR-27a was tested with qRT-PCR from pellets of pulled down assay.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003e \\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eTranswell assay\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eFor transwell invasion assay, cells were plated as described previously. Briefly, the wells were separated into upper and bottom chambers by a membrane with 8 \\u0026micro;m round pores (Corning). The bottom of the upper chamber was coated with extracellular matrix gel mimicking the tumor micro-environment \\u003cem\\u003ein vivo\\u003c/em\\u003e. Cells were plated on the top of the gel in the upper chamber while the media in the bottom chamber was supplemented with chemokines to induce cells penetrate the membrane and grow on the bottom surfaces of membrane. After 48 hrs of treatment, the number of cells growing in the bottom surface of membrane was quantified. \\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eWestern blot analysis\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe protein electrophoreses were conducted on SDS-PAGE gel. Moreover, the blocked membrane transferred with proteins was dealt with skim milk and probed with primary antibody. The primary antibodies were utilized in our work, including \\u0026beta;-actin (13E5) (Cell Signaling, Danvers, MA, USA; CST 4970S, dilution 1: 2,000), FAM172A (Abcam; ab 121364: dilution 1:2,000), anti-GRP78/BiP antibody (ab21685, dilution 1: 800; Abcam, Cambridge, MA, USA); anti-pATF2 (CST 40749, dilution 1: 800; Cell Signalling, Danvers, MA, USA); polyclonal rabbit anti-Human EMP-1 Antibody (aa35-63, LS-B12859, dilution 1: 600; Absolute Biotech, WA, USA). Finally, the chemiluminescence examination was performed based on the secondary HRP antibody using an exposed film. \\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003e \\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eStatistics\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe data were shown as mean \\u0026plusmn; SD (Standard deviation). The differences between the two groups were analyzed with the Student\\u0026rsquo;s \\u003cem\\u003et\\u003c/em\\u003e-test. Kruskal-Wallis ANOVA was used to analyze the abnormally distributed data among groups. SPSS software (version 18.0) was utilized to perform these statistical analyses. \\u003cem\\u003eP\\u003c/em\\u003e\\u0026lt; 0.05 was significant statistically.\\u003c/p\\u003e\\n\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eDifferential miRNAs and lncRNAs expression \\u003c/em\\u003e\\u003c/strong\\u003e\\u003cstrong\\u003e\\u003cem\\u003eanalysis\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe differential expression of miRNAs was further valuated in NPC patients between the tumor tissues and their paired histologically normal tissues adjacent to the tumor (NAT) based on miRNA sequencing. Consequently, there were differentially expressed eleven miRNAs between the two groups, including nine upregulated expression of miRNAs and two downregulated miRNAs in FAM172A deficient mice (\\u003cstrong\\u003eSupplementary Table 2\\u003c/strong\\u003e). Additionally, the relative miR-27a expression in tumor tissues of NPC patients was significantly enhanced than that in the paired normal tissues (\\u003cstrong\\u003eFigure 1A\\u003c/strong\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eMoreover, after a series of screening works on lncRNAs sequencing, the differential lncRNAs expression analysis of the tumor tissues and their paired histologically normal tissues adjacent to the tumor (NAT) in NPC patients were performed, too. We found five lncRNAs, including three upregulated and two downregulated lncRNAs with stable expression differences (\\u003cstrong\\u003eSupplementary Table 3\\u003c/strong\\u003e). \\u003c/p\\u003e\\n\\u003cp\\u003eAfter screening the lengths, adjacent coding genes, and other multiple conditions of the above ten lncRNAs, then these results show that NR_028080.1 had the full-length sequence in the open database. By querying the NCBI database, it was found that there was no report of NR_028080.1. Furthermore, the analysis of biological informatics confirmed that NR_028080.1 was the transcript coming from \\u003cem\\u003eFAM172A \\u003c/em\\u003egene, which was extensively studied by our team. Based on its expression level in tissues of NPC, we named NR_028080.1 as LncRNA-DRNPC (Down-regulated in nasopharyngeal carcinoma). \\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, qRT-PCR experiments were conducted to identify the differential expression of miRNAs and lncRNAs in tissues. These results demonstrated the high expression level of miR-27a (\\u003cstrong\\u003eFig\\u003c/strong\\u003e\\u003cstrong\\u003eure 1A\\u003c/strong\\u003e) and low expression level of LncRNA-DRNPC (\\u003cstrong\\u003eFig\\u003c/strong\\u003e\\u003cstrong\\u003eure 1B\\u003c/strong\\u003e) in tumor tissues coming from NPC patients than that in the control normal tissues. It was in line with the results of sequencing. \\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eLncRNA-DRNPC bound to miR-27a\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWhereafter, two biological informatics software DIANA-LncBase and miRcode, was used to predict the combination between the differential expressed miRNAs and lncRNAs selected from the above sequencing experiments. Then, the results indicated that miR-27a had binding sites of LncRNA-DRNPC (\\u003cstrong\\u003eFigure 2A\\u003c/strong\\u003e). \\u003c/p\\u003e\\n\\u003cp\\u003eMoreover, the overexpression (\\u003cstrong\\u003eFigure 2B\\u003c/strong\\u003e) and knockdown (\\u003cstrong\\u003eFigure 2C\\u003c/strong\\u003e) of LncRNA-DRNPC shifted miR-27a expression level. It suggested the targeted relationship between them, then the assay of RNA pull-down and dual luciferase reporter assay were conducted to confirm their interaction. According to the assay of dual luciferase reporter gene, the mimics of miR-27a and LncRNA-DRNPC-WT were co-transfected into NPC cell line CNE2. These results figured out the significantly decrease of the fluorescence induced by miR-27a (\\u003cstrong\\u003eFigure 2D\\u003c/strong\\u003e). Meanwhile, it identified LncRNA-DRNPC directly targeted miR-27a. Besides, Ago2 was detected in the mixed extraction of cell lines and LncRNA-DRNPC labeled with biotin according to RNA pull-down assay (\\u003cstrong\\u003eFigure 2E\\u003c/strong\\u003e), and it suggested the combination between RISC complex and LncRNA-DRNPC. In addition, the high expression level of miR-27a was found in DRNPC-pulled-down pellets based on qRT-PCR experiment (\\u003cstrong\\u003eFigure 2F\\u003c/strong\\u003e). These results suggested that as a ceRNA, LncRNA-DRNPC could regulate miR-27a.\\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003emiR-27a inhibited FAM172A expression in NPC cell line\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe luciferase reporter gene system was further utilized to prove miR-27a straight target FAM172A in NPC cell line. Our results demonstrated that 3\\u0026rsquo; UTR luciferase activity of FAM172A downregulated clearly in CNE1 cell line after treatment with mimics of miR-27a (\\u003cstrong\\u003eFigure 3A\\u003c/strong\\u003e). Nevertheless, the mutation of FAM172A group did not exhibit significant alteration (\\u003cstrong\\u003eFigure 3B\\u003c/strong\\u003e). Meanwhile, miR-27a did not bind to the 3\\u0026rsquo; UTRs of GRP78/BiP , CHOP, ATF2, and EMP-1 (data not shown). \\u003c/p\\u003e\\n\\u003cp\\u003eMoreover, the role of miR-27a in FAM172A protein expression was explored. The results demonstrated that miR-27a suppressed FAM172A protein expression in CNE1 cell line (\\u003cstrong\\u003eFigure 3C\\u003c/strong\\u003e). In a word, it stated that miR-27a straight regulated the expression of FAM172A mRNA by binding to its 3\\u0026prime;-UTR, and further affected its protein expression level.\\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eLncRNA-DRNPC competed with miR-27a for binding to FAM172A, and participated in the regulation of ER stress-related metastasis \\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThrough examining the expression levels of ER stress marker protein GRP78 and its key signal transduction pathway protein pATF2, we found that the LncRNA-DRNPC resulted in obviously increased expression levels of FAM172A, but the remarkably decreased of GRP78 and pATF2 expression level in CNE2 cell line. On the contrary, miR-27a caused significantly increased expression levels of GRP78 and pATF2 through suppressing FAM172A expression. However, the above biological function of miR-27a could be reversed by LncRNA-DRNPC (\\u003cstrong\\u003eFigure 4A\\u003c/strong\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, the transwell invasion assay was conducted to explore the bio-function of above interaction net. It was declared that LncRNA-DRNPC effectively inhibited cell invasion of NPC cell line CNE2 (\\u003cem\\u003eP\\u003c/em\\u003e\\u0026lt;0.01). On the contrary, miR-27a resulted in a significant promotion of cell invasion of NPC cell line CNE2 (\\u003cem\\u003eP\\u003c/em\\u003e\\u0026lt;0.01). However, the cell invasion induced by miR-27a could be reversed by LncRNA-DRNPC (\\u003cstrong\\u003eFigure 4B and 4C\\u003c/strong\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eThese results indicated that LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the cell invasion associated with ER stress in NPC cell line CNE2.\\u003c/p\\u003e\\n\\n\\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eEBV-LMP1-mediated ER stress-metastasis associated with the interaction net of LncRNA-DRNPC-miR-27a-FAM172A\\u003c/em\\u003e\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, qRT-PCR experiments were conducted to identify the differential expression of miRNAs and lncRNAs in CNE2. These results demonstrated the high expression level of miR-27a (\\u003cstrong\\u003eFig\\u003c/strong\\u003e\\u003cstrong\\u003eure 5A\\u003c/strong\\u003e) and low expression level of LncRNA-DRNPC (\\u003cstrong\\u003eFig\\u003c/strong\\u003e\\u003cstrong\\u003eure 5B\\u003c/strong\\u003e) in the group of treatment with EBV-LMP1. EBV-LMP1 (Latent membrane protein 1) is the major oncoprotein encoded by EBV, and contributes to the metastatic of NPC.\\u003c/p\\u003e\\n\\u003cp\\u003eMoreover，LMP1 protein induced down-expression of FAM172A, over-expression of the ERS marker protein GRP78, and the up-regulated of EMP-1. EMP-1 was the key regulator of tumor invasion (\\u003cstrong\\u003eFig\\u003c/strong\\u003e\\u003cstrong\\u003eure 6A\\u003c/strong\\u003e). Furthermore, the results of transwell invasion assay demonstrated that EBV-LMP1 promoted ERS in NPC cell line CNE2, and induced cell invasion of NPC. However, the cell invasion induced by EBV-LMP1 could be reversed by LncRNA-DRNPC (\\u003cstrong\\u003eFigure 6B and 6C\\u003c/strong\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eThese results indicated that LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the NPC cell invasion associated with ER stress.\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eAlthough platinum-containing induction chemotherapy and radiotherapy is used as the first-line standard treatment, the median progression-free survival of NPC patients after treatment is only about seven months. During tumor metastasis progression, there is no standard method for second-line treatment, and the median overall survival in NPC is only about one year. Additionally, more than 130,000 new cases of NPC were diagnosed worldwide in 2020, nearly half of which occurred in China, especially in southern China [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e]. The incidence and mortality of NPC in China are higher than the global average, and the death cases account for about 40% of all deaths from NPC in the world [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. Therefore, to provide the new treatment strategies for patients with NPC is the crucial significance for improving the prognosis of advanced NPC.\\u003c/p\\u003e \\u003cp\\u003eThe principal oncoprotein of EBV (Epstein-Barr Virus), LMP1 (Latent Membrane Protein 1) is the superfamily receptor of TNF (Tumor Necrosis Factor) [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. The features of LMP1 figure out its constitutive activity, and PRRs (Pathogen Recognition Receptors) functions, such as NRF2 of the master antioxidative transcription factor, IRFs subset. Moreover, its signal transduction lead to the activation of Akt, AP1, and NF-κB, and include the utilization of kinase cascades, adaptors, and TRAFs [\\u003cspan additionalcitationids=\\\"CR13 CR14 CR15 CR16 CR17 CR18 CR19 CR20 CR21\\\" citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e]. Furthermore, LMP1 is a pleiotropic functional factor involved in the balance and reprogram in the broad spectrum of cellular functions, including telomere elongation, immune defenses, extracellular vehicles, DNA damage response, epigenetics, metabolism, ubiquitin machinery [\\u003cspan additionalcitationids=\\\"CR14 CR15 CR16 CR17\\\" citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e]. Functionally, LMP1 promote the development of the tumor microenvironment, metastasis, invasion, cell survival and proliferation, anchorage-independent cell growth, angiogenesis, as well as oncogenic transformation [\\u003cspan additionalcitationids=\\\"CR19 CR20 CR21\\\" citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e]. Therefore, we explored the relationship between LMP1 and metastasis associated with ER stress.\\u003c/p\\u003e \\u003cp\\u003eEMP1 was the key role in metastatic recurrence, then it was utilized in our work to confirm metastasis status [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. At first, the LncRNAs closely related to nasopharyngeal carcinoma was screened by our research group through the next generation sequencing and proved to be the transcript of the tumor suppressor gene \\u003cem\\u003eFAM172A\\u003c/em\\u003e in NPC, so it was named DRNPC. Consequently, the next-genesequencing experiments of noncoding RNAs were conducted based on the NPC tissues of patients. The differential microRNAs and lncRNA were found and identified, including miR-27a and LncRNA-DRNPC. In addition, our results further confirmed that miR-27a had binding sites with LncRNA-DRNPC, and LncRNA-DRNPC could regulate miR-27a as a ceRNA. At the mean time, miR-27a inhibited FAM172A expression in NPC cell line. Moreover, miR-27a resulted in a significant promotion of cell invasion of NPC cell line CNE2. However, the cell invasion induced by miR-27a could be reversed by LncRNA-DRNPC. LncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the cell invasion associated with ER stress in NPC cell line CNE2. Secondly, we found that EBV-LMP1 promoted ERS in NPC cell line CNE2, and induced cell invasion of NPC. However, the cell invasion induced by EBV-LMP1 could be reversed by LncRNA-DRNPC.\\u003c/p\\u003e \\u003cp\\u003eFinally, LncRNA-DRNPC inhibited EBV infection-associated nasopharyngeal carcinoma cell metastasis and endoplasmic reticulum stress, the metastasis and ER stress were positively correlated in NPC. LncRNA-DRNPC can bind to FAM172A, while miR-27a can degrade its target gene FAM172A. Therefore, we speculated that LncRNA-DRNPC might inhibit the progression of the expression of the target gene FAM172A through competitive binding with miR-27a, and reverse the progression of LMP1 encoded by Epstein Barr virus (EBV). The endoplasmic reticulum stress induced by LMP1 can promote the metastasis process of nasopharyngeal carcinoma cells. This project aims to further explore the molecular mechanism of LncRNA-DRNPC's regulation of EBV-LMP1-mediated ER stress-nasopharyngeal carcinoma cell invasion through the construction of over-expression of LncRNA-DRNPC and miR-27a silencing. To elucidate the mechanism of LncRNA-DRNPC as a ceRNA of miR-27a in the occurrence and development of EBV infection-associated nasopharyngeal carcinoma, and to provide key new molecular targets for the prevention and treatment of EBV infection-associated nasopharyngeal carcinoma.\\u003c/p\\u003e\"},{\"header\":\"Conclusion\",\"content\":\"\\u003cp\\u003eOur work illustrated that LncRNA-DRNPC competitively binds miR-27a to inhibit EBV-LMP1-mediated ER (Endoplasmic reticulum) stress-metastasis in nasopharyngeal carcinoma (NPC).\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eEthics approval and consent to participate\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis study was approved by the Medical Research Ethics Committee of the The First People\\u0026rsquo;s Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology. Each patient was gave informed con[1]sent and signed a written informed consent form.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent for publication\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable.\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAvailability of data and materials\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe data used to support the findings of this study are included within the article.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCompeting interests\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare that they have no competing interests.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthors\\u0026apos; contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eXBC, LC, and JQZ contributed to the conception of the study; XBC, and LC contributed significantly to performing these experiments; XBC, LC, THL, CF, and JQZ performed the data analyses and wrote the manuscript; XBC, CL, and JQZ helped perform the analysis with constructive discussions.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis work was supported by grants from the National Natural Science Foundation of China (No. 82360224),the Open Project of Yunnan Provincial Key Laboratory of Clinical Virology (202205AG070053), the Key Basic Research Program of Yunnan Province (No. 202201AS070065), and Kunming University of Science and Technology Medical Joint Project (KUST-KH2022038Y). The study sponsors had no involvement in the work.\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eSiegel RL, Miller KD, Fuchs HE, Jemal A, Cancer Statistics (2021) CA Cancer J Clin 2021; 71: 7\\u0026ndash;33\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChen YP, Chan ATC, Le QT, Blanchard P, Sun Y, Ma J (2019) Nasopharyngeal carcinoma. Lancet 394:64\\u0026ndash;80\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNiedobitek G (2000) Epstein-Barr virus infection in the pathogenesis of nasopharyngeal carcinoma. Mol Pathol 53:248\\u0026ndash;254\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZheng ZQ, Li ZX, Zhou GQ, Lin L, Zhang LL, Lv JW et al (2019) Long Noncoding RNA FAM225A Promotes Nasopharyngeal Carcinoma Tumorigenesis and Metastasis by Acting as ceRNA to Sponge miR-590-3p/miR-1275 and Upregulate ITGB3. Cancer Res 79:4612\\u0026ndash;4626\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWang L, Xu T, Cui X, Han M, Zhou LH, Wei ZX et al (2020) Downregulation of lncRNA SNHG7 inhibits proliferation and invasion of nasopharyngeal carcinoma cells through repressing ROCK1. Eur Rev Med Pharmacol Sci 24:7917\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCubillos-Ruiz JR, Bettigole SE, Glimcher LH (2017) Tumorigenic and Immunosuppressive Effects of Endoplasmic Reticulum Stress in Cancer. Cell 168:692\\u0026ndash;706\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLiu S, Li Y, Li Z (2022) Salidroside suppresses the activation of nasopharyngeal carcinoma cells via targeting miR-4262/GRP78 axis. Cell Cycle 21:720\\u0026ndash;729\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHe B (2006) Viruses, endoplasmic reticulum stress, and interferon responses. Cell Death Differ 13:393\\u0026ndash;403\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eShen WF, Feng ZQ, Wang P, Zhang JQ (2017) FAM172A controls endoplasmic reticulum (ER) stress related to NF-κB signaling pathway in hepatocellular carcinoma. RSC Adv 7:51870\\u0026ndash;51878\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003ePertea M, Kim D, Pertea GM, Leek JT, Salzberg SL (2016) Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. Nat Protoc 11:1650\\u0026ndash;1667\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKozomara A, Griffiths-Jones S (2011) miRBase: integrating microRNA annotation anddeep-sequencing data. Nucleic Acids Res 39:152\\u0026ndash;157\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWang L, Ning S (2021) New Look of EBV LMP1 Signaling Landscape. Cancers (Basel) 13:5451\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eAwasthi P, Dwivedi M, Kumar D, Hasan S (2023) Insights into intricacies of the Latent Membrane Protein-1 (LMP-1) in EBV-associated cancers. Life Sci 313:121261\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMunz C (2019) Latency and lytic replication in Epstein-Barr virus-associated oncogenesis. Nat Rev Microbiol 17:691\\u0026ndash;700\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChen J, Sathiyamoorthy K, Zhang X, Schaller S, Perez White BE, Jardetzky TS, Longnecker R (2018) Ephrin receptor A2 is a functional entry receptor for Epstein-Barr virus. Nat Microbiol 3:172\\u0026ndash;180\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhang H, Li Y, Wang HB, Zhang A, Chen ML, Fang ZX et al (2018) Ephrin receptor A2 is an epithelial cell receptor for Epstein-Barr virus entry. Nat Microbiol 3:164\\u0026ndash;171\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCheerathodi MR, Meckes DG (2018) The Epstein-Barr virus LMP1 interactome: Biological implications and therapeutic targets. Future Virol 13:863\\u0026ndash;887\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWang LW, Jiang S, Gewurz BE (2017) Epstein-Barr Virus LMP1-Mediated Oncogenicity. J Virol 91:e01718\\u0026ndash;e01716\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eDawson CW, Port RJ, Young LS (2012) The role of the EBV-encoded latent membrane proteins LMP1 and LMP2 in the pathogenesis of nasopharyngeal carcinoma (NPC) Semin. Cancer Biol 22:144\\u0026ndash;153\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMorris M, Dawson C, Young L (2009) Role of the Epstein-Barr virus-encoded latent membrane protein-1, LMP1, in the pathogenesis of nasopharyngeal carcinoma. Future Oncol 5:811\\u0026ndash;825\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLavorgna A, Harhaj EW (2012) EBV LMP1: New and shared pathways to NF-kappaB activation. Proc Natl Acad Sci USA 109:2188\\u0026ndash;2189\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eIkeda O, Miyasaka Y, Yoshida R, Mizushima A, Oritani K, Sekine Y et al (2010) BS69 cooperates with TRAF3 in the regulation of Epstein-Barr virus-derived LMP1/CTAR1-induced NF-κB activation. FEBS Lett 584:865\\u0026ndash;872\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCa\\u0026ntilde;ellas-Socias A, Cortina C, Hernando-Momblona X, Palomo-Ponce S, Mulholland EJ, Turon G, Mateo L et al (2022) Metastatic recurrence in colorectal cancer arises from residual EMP1\\u0026thinsp;+\\u0026thinsp;cells. Nature ; 611): 603\\u0026ndash;613\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":true,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true},\"keywords\":\"DRNPC (Down-regulated in nasopharyngeal carcinoma), FAM172A gene, ER (Endoplasmic reticulum) stress, metastasis, EBV (Epstein Barr virus), LMP1 (Latentmembraneprotein-1)\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-5330415/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-5330415/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003ch2\\u003eBackground\\u003c/h2\\u003e \\u003cp\\u003eThe details of the regulatory mechanism in metastasis of nasopharyngeal carcinoma related to lncRNAs are unclear.\\u003c/p\\u003e\\u003ch2\\u003eMethods\\u003c/h2\\u003e \\u003cp\\u003eThe bio-functions of LncRNA-DRNPC-miR-27a-FAM172A interaction net in the relationships between ER stress, NPC invasion were investigated.\\u003c/p\\u003e\\u003ch2\\u003eResults\\u003c/h2\\u003e \\u003cp\\u003eThere was high expression of miR-27a and low expression of LncRNA-DRNPC in NPC tumor tissues. LncRNA-DRNPC regulated miR-27a as a ceRNA, and miR-27a inhibited FAM172A expression, which participated in the regulation of ER stress-related NPC invasion induced by EBV-LMP1.\\u003c/p\\u003e\\u003ch2\\u003eConclusion\\u003c/h2\\u003e \\u003cp\\u003eLncRNA-DRNPC competes with miR-27a mediated by FAM172A and participated in the regulation of the NPC cell invasion associated with ER stress.\\u003c/p\\u003e\",\"manuscriptTitle\":\"LncRNA-DRNPC competitively binds miR-27a to inhibit EBV-LMP1-mediated ER (Endoplasmic reticulum) stress-metastasis in nasopharyngeal carcinoma (NPC)\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-11-06 11:03:14\",\"doi\":\"10.21203/rs.3.rs-5330415/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"cbbfd042-a236-4ce4-ba95-a374e349a277\",\"owner\":[],\"postedDate\":\"November 6th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2025-02-05T04:08:17+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2024-11-06 11:03:14\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-5330415\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-5330415\",\"identity\":\"rs-5330415\",\"version\":[\"v1\"]},\"buildId\":\"8U1c8b4HqxoKbykW_rLl7\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}