Knock Down of LINC00504 Represses Proliferation and Invasion via Regulation of miR-140-5p in Breast Cancer

Knock Down of LINC00504 Represses Proliferation and Invasion via Regulation of miR-140-5p 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 Help Center Sign In Submit a Preprint Cite Share Download PDF Research Knock Down of LINC00504 Represses Proliferation and Invasion via Regulation of miR-140-5p in Breast Cancer tieying hou, long ye, qingsong qin, shulin wu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-38135/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 : Breast cancer is one of the most common cancer in the world. Emerging evidence has demonstrated the critical role of long noncoding RNAs (lncRNAs) in the development of breast cancer. In this study, we aimed to investigate the role of LINC00504 in breast cancer progression. Methods : Quantification real-time PCR was used to analyzed the expression levels of LINC00504 and miR‐140-5p in breast cancer tissues and cell lines. Cell proliferation, migration and invasion were assessed by Cell Counting Kit‐8, transwell assay and Immunofluorescence. Dual-luciferase reporter assay and RNA Immunoprecipitation assay were performed to verify the interaction between LINC00504 and miR‐140-5p. The expression levels of VEGFA, CDH1 and VIM were demonstrated by western blot assays. Result : Here, we found that LINC00504 is up regulated in breast cancer tissues and cell lines. Down regulation of LINC00504 mediated by shRNA suppressed the proliferation, migration, and invasion of breast cancer cells in vitro and in vivo. Furthermore, LINC00504 was found to competitively regulate miR‐140-5p via targeting VEGFA. Inhibition of miR‐140-5p attenuated the knockdown-LINC00504 induced inhibition of breast cancer cell proliferation and invasion. Conclusion : Taken together, our results demonstrated the mechanism of the LINC00504–miR‐140-5p–VEGFA axis in breast cancer cell proliferation and invasion and may lead to new lncRNA-based diagnostics or therapeutics for breast cancer. Cell Communication and Signaling breast cancer LINC00504 proliferation invasion miR-140-5p VEGFA Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Full Text Supplementary Files CoverLetter.doc CoverLetter.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. 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Also discoverable on Platform About Our Team In Review Editorial Policies 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-38135","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":736929,"identity":"0922ec13-f48e-4883-85b6-ad4922817262","order_by":0,"name":"tieying hou","email":"","orcid":"","institution":"Shantou University Medical College","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"tieying","middleName":"","lastName":"hou","suffix":""},{"id":736930,"identity":"5fe06252-7c7b-4023-82f8-4338a482d297","order_by":1,"name":"long ye","email":"","orcid":"","institution":"Guangdong Provincial People's Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"long","middleName":"","lastName":"ye","suffix":""},{"id":736931,"identity":"d6bd0457-0af5-4b87-a6c5-87b547be41fb","order_by":2,"name":"qingsong qin","email":"","orcid":"","institution":"Shantou University Medical College","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"qingsong","middleName":"","lastName":"qin","suffix":""},{"id":736932,"identity":"60ec2ec4-01d3-4a2c-bba1-c54ea4158817","order_by":3,"name":"shulin wu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzklEQVRIiWNgGAWjYDACCQbGAwlAmp+9+cCBDxXEaWEAa5HsOZZ4cMYZYrWAaIMZPsaHeVuI0CE/u8fgwMMdhxM3SPB8OMDbwCDPL3YAvxaDO2cMDiSeOZy4Xbp3wwHJHQyGM2cnENAikQPU0nY7ceecsxsOGJ5hSDC4TUCL/Ayolg03ch4AGURoYbiB0MJw4CAxWgxupBUAtfw3ntlzzOBgwxkJwn6Rn5G88eHPtjTZfvbmx5//VNjI80sTchgakCBN+SgYBaNgFIwC7AAALkVUYRyCVc8AAAAASUVORK5CYII=","orcid":"","institution":"","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"shulin","middleName":"","lastName":"wu","suffix":""}],"badges":[],"createdAt":"2020-06-27 16:13:51","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-38135/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-38135/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":1460540,"identity":"5d0cfb93-c74b-4978-a143-d199a1e5ce25","added_by":"auto","created_at":"2020-06-30 18:21:14","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":607923,"visible":true,"origin":"","legend":"Figure1. LINC00504 was up regulated in breast cancer tissues and cells. The expression of\nLINC00504 in breast cancer patients and healthy people was assessed in The Cancer Genome Atlas\n(TCGA) database(A). qRT-PCR assay was used to assess the expression of LINC00504 in 28 pairs\nbreast cancer tissues and adjacent noncancerous tissues(B), breast cancer cell lines (BT549, T47D,\nMCF-7, SKBR3 and MDA-MB-231) and normal breast cell line MCF-10A(C). **P\u003c0.01 vs normal\ntissues or MCF-10A.\n\n","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/Figure1.jpg"},{"id":1460541,"identity":"e238e49c-8bea-4c13-99a7-4637f1e61638","added_by":"auto","created_at":"2020-06-30 18:21:14","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1231266,"visible":true,"origin":"","legend":"The effect of LINC00504 knockdown on the proliferation, cell cycle distribution of breast\ncancer cells in vitro. SKBR3 and MDA-MB-231 cells were transfected with sh-control or sh-\nLINC00504. LINC00504 expression was detected by qRT-PCR when transfected in the two cells(A).\ntransfected cells proliferation was detected by CCK-8 and colony formation assays(B-D). Flow\ncytometer was used to analyze the transfected cells at different cell cycle phase (G1, S, and G2)(E).\n**P\u003c0.01 vs sh-control.\n","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/Figure2.jpg"},{"id":1460542,"identity":"d6fe2f62-0f47-4b0b-9b17-20f2d1fbf8ec","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":571596,"visible":true,"origin":"","legend":"The effect of LINC00504 knockdown on the proliferation in vivo. Transfected cells were\nplanted in mice, and tumor growth was measured. **P\u003c0.01 vs sh-control.\n","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/Figure3.jpg"},{"id":1460543,"identity":"85d1d935-60a1-4f89-9e63-4871c338d251","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2800574,"visible":true,"origin":"","legend":"The influence of LINC00504 knockdown on invasion, and EMT of breast cancer cells.\nTransfected cells migration and invasion capacities were assessed by transwell assays(A-B).\nImmunofluorescence was used to detect the variety of VIM and CDH1 in Transfected cells.\n**P\u003c0.01 vs sh-control.\n","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/Figure4.jpg"},{"id":1460544,"identity":"073e4e3e-affd-49a6-8b0b-74849c8dcfb7","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1120898,"visible":true,"origin":"","legend":"LINC00504 directly binded to miR-140-5p in SKBR3 and MDA-MB-231 cells. The\npotential binding sites between LINC00504 and miR-140-5p, and the mutant in the seed region(A).\nThe expression of miR-140-5p was detected in sh-control or sh-LINC00504 transfected cells(B).\nDual-luciferase reporter assays were performed by transfecting with LINC00504-WT or\nLINC00504-Mut constructs into SKBR3 and MDA-MB-231 cells with miR-NC mimics or miR-\n140-5p(C). SKBR3 and MDA-MB-231 cells were transfected with miR-NC mimics or miR-140-5p\nmimics, followed by the measurement of LINC00504 mRNA enrichment with anti-Ago2 by qRT-\nPCR, and anti IgG served as control(D,E). Relationship between levels of LINC00504 and miR-\n140-5p in breast cancer tissues(F). **P\u003c0.01 vs sh-control or miR-NC.\n","description":"","filename":"Figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/Figure5.jpg"},{"id":1460545,"identity":"c6f0ef89-46f3-4357-9ede-f66efeb2c265","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":528257,"visible":true,"origin":"","legend":"LINC00504 promoted the migration, invasion, and EMT of breast cancer cells, and it was\nregulated by miR-140-5p-VEGFA axis. SKBR3 and MDA-MB-231 cells were transfected with sh-\ncontrol, sh-LINC00504 or sh-LINC00504+miR-140-5p inhibitor, followed by the trial of colony\nformation for cell proliferation(A), transwell assays for cell migration(B) and cell invasion(C),\nwestern blot analysis for VEGFA, CDH1, VIM levels(D). **P\u003c0.01 vs corresponding control.\n","description":"","filename":"Figure6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/Figure6.jpg"},{"id":13512390,"identity":"49bb81c2-6db2-4e48-a0e0-c6b490edf175","added_by":"auto","created_at":"2021-09-16 23:55:36","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":871537,"visible":true,"origin":"","legend":"","description":"","filename":"manuscriptnew4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1_covered.pdf"},{"id":1460548,"identity":"e046a3d1-0f71-4db5-a2b7-3706faee0a62","added_by":"auto","created_at":"2020-06-30 18:21:16","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":267679,"visible":true,"origin":"","legend":"","description":"","filename":"manuscriptnew4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1_stamped.pdf"},{"id":1460538,"identity":"97af0557-4c11-49f5-b099-fd2e36744f8a","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":238906,"visible":true,"origin":"","legend":"","description":"","filename":"manuscriptnew4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/manuscriptnew4.pdf"},{"id":1460546,"identity":"39dd0c68-9a92-4220-9c2d-80ec00f47e2c","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":238906,"visible":true,"origin":"","legend":"","description":"","filename":"manuscriptnew4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/manuscriptnew4.pdf"},{"id":1460547,"identity":"faba4850-c4c9-4c3d-9012-119d6e161d70","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"doc","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":26624,"visible":true,"origin":"","legend":"","description":"","filename":"CoverLetter.doc","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/CoverLetter.doc"},{"id":1460539,"identity":"53c880cc-533b-49d2-8979-129efd88dc3e","added_by":"auto","created_at":"2020-06-30 18:21:15","extension":"doc","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":26624,"visible":true,"origin":"","legend":"","description":"","filename":"CoverLetter.doc","url":"https://assets-eu.researchsquare.com/files/rs-38135/v1/CoverLetter.doc"}],"financialInterests":"","formattedTitle":"\u003cp\u003eKnock Down of LINC00504 Represses Proliferation and Invasion via Regulation of miR-140-5p in Breast Cancer\u003c/p\u003e","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-38135/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"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":false,"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":"breast cancer, LINC00504, proliferation, invasion, miR-140-5p, VEGFA","lastPublishedDoi":"10.21203/rs.3.rs-38135/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-38135/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: Breast cancer is one of the most common cancer in the world. Emerging evidence has demonstrated the critical role of long noncoding RNAs (lncRNAs) in the development of breast cancer. In this study, we aimed to investigate the role of LINC00504 in breast cancer progression. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: Quantification real-time PCR was used to analyzed the expression levels of LINC00504 and miR‐140-5p in breast cancer tissues and cell lines. Cell proliferation, migration and invasion were assessed by Cell Counting Kit‐8, transwell assay and Immunofluorescence. Dual-luciferase reporter assay and RNA Immunoprecipitation assay were performed to verify the interaction between LINC00504 and miR‐140-5p. The expression levels of VEGFA, CDH1 and VIM were demonstrated by western blot assays. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResult\u003c/strong\u003e: Here, we found that LINC00504 is up regulated in breast cancer tissues and cell lines. Down regulation of LINC00504 mediated by shRNA suppressed the proliferation, migration, and invasion of breast cancer cells in vitro and in vivo. Furthermore, LINC00504 was found to competitively regulate miR‐140-5p via targeting VEGFA. Inhibition of miR‐140-5p attenuated the knockdown-LINC00504 induced inhibition of breast cancer cell proliferation and invasion. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: Taken together, our results demonstrated the mechanism of the LINC00504–miR‐140-5p–VEGFA axis in breast cancer cell proliferation and invasion and may lead to new lncRNA-based diagnostics or therapeutics for breast cancer.\u003c/p\u003e","manuscriptTitle":"Knock Down of LINC00504 Represses Proliferation and Invasion via Regulation of miR-140-5p in Breast Cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-06-30 18:21:13","doi":"10.21203/rs.3.rs-38135/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":"a37f3acc-a587-442f-a018-45bf0384213a","owner":[],"postedDate":"June 30th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":136298,"name":"Cell Communication and Signaling"}],"tags":[],"updatedAt":"2020-06-30T18:21:14+00:00","versionOfRecord":[],"versionCreatedAt":"2020-06-30 18:21:13","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-38135","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-38135","identity":"rs-38135","version":["v1"]},"buildId":"GqpaHPwrfC8PjnIFayRh5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}