Detachment-activated FAK-STAT3-NNMT inhibits anoikis by enhancing fatty acid oxidation in breast cancer

Detachment-activated FAK-STAT3-NNMT inhibits anoikis by enhancing fatty acid oxidation in breast cancer | 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 Detachment-activated FAK-STAT3-NNMT inhibits anoikis by enhancing fatty acid oxidation in breast cancer Qingchao Tong, Yilei Ma, Yuzhen Gao, Lin Zhou, Qiang Fang, Sining Fang, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3933796/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 Purpose: Breast cancer metastasis claims the majority of breast cancer-related deaths. Anoikis resistance is a key prerequisite for tumor metastasis. Possible mechanism of evading anoikis in breast cancer remains unclear. Our previous research demonstrated that Nicotinamide N-methyltransferase (NNMT) has essential functions in cancer development and metastasis. However, the spercific functional mechanisms of NNMT in breast cancer anoikisp remain to be explored Method: Immunofluorescence staining, confocal microscopy analysis, immunoblotting and qPCR were used to analyze NNMT expression. Flow cytometry and soft agar assay were used to analyze the proportion of breast cancer cells undergoing anoikis. ChIP assay and detached cell model were used to confirm the presence of FAK-STAT3-NNMT in detached breast cancer cells. BODIPY staining was used to visualize lipid deposit in the cytoplasm, and Seahorse analysis was used to directly measure FAO rate of attached and detached breast cancer cells. Results: Detachment of breast cancer cells activates the FAK-STAT3 axis to increase the transcription of NNMT, which then protects breast cancer cells from anoikis by enhancing fatty acid oxidation (FAO). Mechanistically, NNMT promotes the expression of CPT1A, a FAO rate-limiting gene, by suppressing PP2A methylation. Furthermore, NNMT-induced FAO accelerates the clearance of reactive oxygen species (ROS), which triggers anoikis and hinders cancer metastasis, by maintaining NADP+/NADPH balance. In vivo experiments show that NNMT-knockdown breast cancer cells colonize the lung much less than that of NNMT high expression cells. Additionally, inhibitors of NNMT or FAO suppressed breast cancer cell metastasis, suggesting that NNMT-mediated FAO enhances the metastatic potential of breast cancer. Conclusion: Our study identifies that NNMT, an enzyme upregulated during detachment via the FAK-STAT3 axis, activates the FAO pathway to resist anoikis of breast cancer cells. anoikis breast cancer NNMT FAO Full Text Additional Declarations No competing interests reported. Supplementary Files SupTables.xlsx supplementaryfigures.docx 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-3933796","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":271868410,"identity":"dd27e21d-6143-40e4-bf1a-af414d7079dc","order_by":0,"name":"Qingchao Tong","email":"","orcid":"","institution":"Zhejiang University School of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Qingchao","middleName":"","lastName":"Tong","suffix":""},{"id":271868411,"identity":"dea009eb-2401-49d0-9304-a460ec30e0c2","order_by":1,"name":"Yilei Ma","email":"","orcid":"","institution":"Sir Run Run Shaw 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cancer","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":"anoikis, breast cancer, NNMT, FAO","lastPublishedDoi":"10.21203/rs.3.rs-3933796/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3933796/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePurpose: Breast cancer metastasis claims the majority of breast cancer-related deaths. Anoikis resistance is a key prerequisite for tumor metastasis. Possible mechanism of evading anoikis in breast cancer remains unclear. Our previous research demonstrated that Nicotinamide N-methyltransferase (NNMT) has essential functions in cancer development and metastasis. However, the spercific functional mechanisms of NNMT in breast cancer anoikisp remain to be explored\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMethod: Immunofluorescence staining, confocal microscopy analysis, immunoblotting and qPCR were used to analyze NNMT expression. Flow cytometry and soft agar assay were used to analyze the proportion of breast cancer cells undergoing anoikis. ChIP assay and detached cell model were used to confirm the presence of FAK-STAT3-NNMT in detached breast cancer cells. BODIPY staining was used to visualize lipid deposit in the cytoplasm, and Seahorse analysis was used to directly measure FAO rate of attached and detached breast cancer cells.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults: Detachment of breast cancer cells activates the FAK-STAT3 axis to increase the transcription of NNMT, which then protects breast cancer cells from anoikis by enhancing fatty acid oxidation (FAO). Mechanistically, NNMT promotes the expression of CPT1A, a FAO rate-limiting gene, by suppressing PP2A methylation. Furthermore, NNMT-induced FAO accelerates the clearance of reactive oxygen species (ROS), which triggers anoikis and hinders cancer metastasis, by maintaining NADP+/NADPH balance. In vivo experiments show that NNMT-knockdown breast cancer cells colonize the lung much less than that of NNMT high expression cells. Additionally, inhibitors of NNMT or FAO suppressed breast cancer cell metastasis, suggesting that NNMT-mediated FAO enhances the metastatic potential of breast cancer.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConclusion: Our study identifies that NNMT, an enzyme upregulated during detachment via the FAK-STAT3 axis, activates the FAO pathway to resist anoikis of breast cancer cells.\u003c/p\u003e","manuscriptTitle":"Detachment-activated FAK-STAT3-NNMT inhibits anoikis by enhancing fatty acid oxidation in breast cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-02-12 06:54:04","doi":"10.21203/rs.3.rs-3933796/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":"8daab7ca-062f-49b7-87a6-ea61ba809604","owner":[],"postedDate":"February 12th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-02-12T09:49:26+00:00","versionOfRecord":[],"versionCreatedAt":"2024-02-12 06:54:04","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3933796","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3933796","identity":"rs-3933796","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}