HNRNPC-Mediated m6A Methylation of ZNF146 Promotes Triple-Negative Breast Cancer Progression through Regulating MRPL21

HNRNPC-Mediated m6A Methylation of ZNF146 Promotes Triple-Negative Breast Cancer Progression through Regulating MRPL21 | 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 HNRNPC-Mediated m6A Methylation of ZNF146 Promotes Triple-Negative Breast Cancer Progression through Regulating MRPL21 Bing Liu, Jiade Li, Huizhe Zhang, Wang Xie, Mengru Cao, Man Liu, and 9 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6299315/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 Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer. The aim of this study was to investigate the expression pattern of ZNF146 in TNBC and explore its role and mechanism in promoting TNBC growth and metastasis. Methods The expression pattern and prognostic value of ZNF146 in breast cancer and normal breast tissue were analyzed using data from TCGA and CPTAC. Lentiviral vectors were used to overexpress or knockdown ZNF146 in TNBC cell lines, which was verified by real-time PCR and western Blot. Functional assays including MTT, EdU, and transwell were performed to investigate the effects of ZNF146 on TNBC cell proliferation, migration, and invasion. Xenograft and tail vein metastasis mouse models were used to evaluate the role of ZNF146 in vivo. The regulatory mechanism of ZNF146 was explored through RNA-Seq, RIP, ChIP, and dual luciferase reporter assays. Results ZNF146 was highly expressed in TNBC and correlated with poor patient survival. Overexpression of ZNF146 promoted TNBC cell proliferation and metastasis both in vitro and in vivo, while knockdown exerted the opposite effect. RNA-Seq analysis revealed MRPL21 as a potential downstream target of ZNF146, which was proved to be an oncogene. ChIP and dual luciferase reporter assays demonstrated that ZNF146 directly binds to the MRPL21 promoter and regulates its expression. MRPL21 overexpression mimicked ZNF146 effects, while its knockdown inhibited TNBC cell proliferation and migration. In addition, HNRNPC, an m6A reader protein, regulates ZNF146 expression in an m6A-dependent manner. Conclusions Our study identifies ZNF146 as a novel oncogenic driver in TNBC, which promotes cancer progression by upregulating MRPL21 expression in an HNRNPC-mediated m6A-dependent manner. These findings provide new insights into the molecular mechanisms underlying TNBC progression and identify potential therapeutic targets. Triple-negative breast cancer ZNF146 m6A methylation HNRNPC MRPL21 Mitochondrial function Full Text Additional Declarations No competing interests reported. Supplementary Files Gel.pptx 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-6299315","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":444394087,"identity":"759cb275-bea6-4fcb-b9f6-8c014e8eb013","order_by":0,"name":"Bing Liu","email":"","orcid":"","institution":"The First Affiliated Hospital of Harbin Medical University","correspondingAuthor":false,"prefix":"","firstName":"Bing","middleName":"","lastName":"Liu","suffix":""},{"id":444394088,"identity":"4b66c97a-de47-4e79-a63b-5158696f9bb7","order_by":1,"name":"Jiade Li","email":"","orcid":"","institution":"Harbin Medical 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MRPL21","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","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":"Triple-negative breast cancer, ZNF146, m6A methylation, HNRNPC, MRPL21, Mitochondrial function","lastPublishedDoi":"10.21203/rs.3.rs-6299315/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6299315/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eTriple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer. The aim of this study was to investigate the expression pattern of ZNF146 in TNBC and explore its role and mechanism in promoting TNBC growth and metastasis.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eThe expression pattern and prognostic value of ZNF146 in breast cancer and normal breast tissue were analyzed using data from TCGA and CPTAC. Lentiviral vectors were used to overexpress or knockdown ZNF146 in TNBC cell lines, which was verified by real-time PCR and western Blot. Functional assays including MTT, EdU, and transwell were performed to investigate the effects of ZNF146 on TNBC cell proliferation, migration, and invasion. Xenograft and tail vein metastasis mouse models were used to evaluate the role of ZNF146 in vivo. The regulatory mechanism of ZNF146 was explored through RNA-Seq, RIP, ChIP, and dual luciferase reporter assays.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eZNF146 was highly expressed in TNBC and correlated with poor patient survival. Overexpression of ZNF146 promoted TNBC cell proliferation and metastasis both in vitro and in vivo, while knockdown exerted the opposite effect. RNA-Seq analysis revealed MRPL21 as a potential downstream target of ZNF146, which was proved to be an oncogene. ChIP and dual luciferase reporter assays demonstrated that ZNF146 directly binds to the MRPL21 promoter and regulates its expression. MRPL21 overexpression mimicked ZNF146 effects, while its knockdown inhibited TNBC cell proliferation and migration. In addition, HNRNPC, an m6A reader protein, regulates ZNF146 expression in an m6A-dependent manner.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eOur study identifies ZNF146 as a novel oncogenic driver in TNBC, which promotes cancer progression by upregulating MRPL21 expression in an HNRNPC-mediated m6A-dependent manner. These findings provide new insights into the molecular mechanisms underlying TNBC progression and identify potential therapeutic targets.\u003c/p\u003e","manuscriptTitle":"HNRNPC-Mediated m6A Methylation of ZNF146 Promotes Triple-Negative Breast Cancer Progression through Regulating MRPL21","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-25 17:08:13","doi":"10.21203/rs.3.rs-6299315/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","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":"da37b479-ec5c-4776-87ce-421875c28cbf","owner":[],"postedDate":"April 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-26T16:53:30+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-25 17:08:13","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6299315","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6299315","identity":"rs-6299315","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}