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<!DOCTYPE html> <html lang="en" dir="ltr" xmlns="http://www.w3.org/1999/xhtml" xmlns:mml="http://www.w3.org/1998/Math/MathML"> <head prefix="og: http://ogp.me/ns# article: http://ogp.me/ns/article# book: http://ogp.me/ns/book#" >  <link rel="dns-prefetch" href="//d33xdlntwy0kbs.cloudfront.net" /> <link rel="dns-prefetch" href="//www.google.com" /> <link rel="dns-prefetch" href="//scholar.google.com" /> <link rel="dns-prefetch" href="//www.googletagmanager.com" /> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <link rel="shortcut icon" href="https://www.biorxiv.org/sites/default/files/images/favicon.ico" type="image/vnd.microsoft.icon" /> <meta name="viewport" content="width=device-width, initial-scale=1" /> <link rel="alternate" type="application/pdf" title="Full Text (PDF)" href="/content/10.1101/2024.11.13.623301v1.full.pdf" /> <link rel="alternate" type="text/plain" title="Full Text (Plain)" href="/content/10.1101/2024.11.13.623301v1.full.txt" /> <meta name="type" content="article" /> <meta name="category" content="article" /> <meta name="HW.identifier" content="/biorxiv/early/2024/11/15/2024.11.13.623301.atom" /> <meta name="HW.pisa" content="biorxiv;2024.11.13.623301v1" /> <meta name="DC.Format" content="text/html" /> <meta name="DC.Language" content="en" /> <meta name="DC.Title" content="A single cysteine residue in vimentin regulates long non-coding RNA XIST to suppress epithelial-mesenchymal transition and stemness in breast cancer" /> <meta name="DC.Identifier" content="10.1101/2024.11.13.623301" /> <meta name="DC.Date" content="2024-11-15" /> <meta name="DC.Publisher" content="Cold Spring Harbor Laboratory" /> <meta name="DC.Rights" content="漏 2024, Posted by Cold Spring Harbor Laboratory. This pre-print is available under a Creative Commons License (Attribution 4.0 International), CC BY 4.0, as described at http://creativecommons.org/licenses/by/4.0/" /> <meta name="DC.AccessRights" content="restricted" /> <meta name="DC.Description" content="Vimentin is a type III intermediate filament (IF) protein, that is induced in a large number of solid tumours. A single cysteine at position 328 in vimentin plays a crucial role in assembly, organisation and stability of IFs. However, its exact function during epithelial mesenchymal transition (EMT) and cancer progression has not been investigated. To investigate this, we have transduced wildtype (WT) and C328S vimentin separately in MCF-7 cells that lack endogenous vimentin. The expression of C328-VIM impacted vimentin-actin interactions and induced EMT-like features that include enhanced cell proliferation, migration, invasion accompanied by reduced cell adhesion when compared to the wildtype cells. Functional transcriptomic studies confirmed the upregulation of EMT and mesenchymal markers, downregulation of epithelial markers as well as acquisition of signatures associated with cancer stemness ( CD56, Oct4, PROCR and CD49f ) thus transforming MCF-7 cells from oestrogen positive to triple reduced ( ESR1, PGR, and HER2 ) status. We also observed a stark increase in the expression of long non-coding RNA, XIST in MCF-7 cells expressing C328-VIM. Targeting the mutant vimentin or XIST by RNA interference partially reversed the phenotypes in C328-VIM expressing MCF-7 cells. Furthermore, introduction of C328-VIM cells into nude mice promoted tumour growth by increasing cancer stemness in an oestrogen independent manner. Altogether, our studies provide insight into how cysteine 328 in vimentin dictates mechano-transduction signals to remodel actin cytoskeleton and protect against EMT and cancer growth via modulating lncRNA XIST . Therefore, targeting vimentin and/or XIST via RNA interference should be a promising therapeutic strategy for breast cancer treatment. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="DC.Contributor" content="Saima Usman" /> <meta name="DC.Contributor" content="W. Andrew Yeudall" /> <meta name="DC.Contributor" content="Muy-Teck Teh" /> <meta name="DC.Contributor" content="Fatemah Ghloum" /> <meta name="DC.Contributor" content="Hemanth Tummala" /> <meta name="DC.Contributor" content="Ahmad Waseem" /> <meta name="article:published_time" content="2024-11-15" /> <meta name="article:section" content="New Results" /> <meta name="citation_title" content="A single cysteine residue in vimentin regulates long non-coding RNA XIST to suppress epithelial-mesenchymal transition and stemness in breast cancer" /> <meta name="citation_abstract" lang="en" content="<p>Vimentin is a type III intermediate filament (IF) protein, that is induced in a large number of solid tumours. A single cysteine at position 328 in vimentin plays a crucial role in assembly, organisation and stability of IFs. However, its exact function during epithelial mesenchymal transition (EMT) and cancer progression has not been investigated. To investigate this, we have transduced wildtype (WT) and C328S vimentin separately in MCF-7 cells that lack endogenous vimentin. The expression of C328-VIM impacted vimentin-actin interactions and induced EMT-like features that include enhanced cell proliferation, migration, invasion accompanied by reduced cell adhesion when compared to the wildtype cells. Functional transcriptomic studies confirmed the upregulation of EMT and mesenchymal markers, downregulation of epithelial markers as well as acquisition of signatures associated with cancer stemness (<i>CD56, Oct4, PROCR and CD49f</i>) thus transforming MCF-7 cells from oestrogen positive to triple reduced (<i>ESR1, PGR, and HER2</i>) status. We also observed a stark increase in the expression of long non-coding RNA, <i>XIST</i> in MCF-7 cells expressing C328-VIM. Targeting the mutant vimentin or <i>XIST</i> by RNA interference partially reversed the phenotypes in C328-VIM expressing MCF-7 cells. Furthermore, introduction of C328-VIM cells into nude mice promoted tumour growth by increasing cancer stemness in an oestrogen independent manner. Altogether, our studies provide insight into how cysteine 328 in vimentin dictates mechano-transduction signals to remodel actin cytoskeleton and protect against EMT and cancer growth via modulating lncRNA <i>XIST</i>. Therefore, targeting vimentin and/or <i>XIST</i> via RNA interference should be a promising therapeutic strategy for breast cancer treatment.</p>" /> <meta name="citation_journal_title" content="bioRxiv" /> <meta name="citation_publisher" content="Cold Spring Harbor Laboratory" /> <meta name="citation_publication_date" content="2024/01/01" /> <meta name="citation_mjid" content="biorxiv;2024.11.13.623301v1" /> <meta name="citation_id" content="2024.11.13.623301v1" /> <meta name="citation_public_url" content="https://www.biorxiv.org/content/10.1101/2024.11.13.623301v1" /> <meta name="citation_abstract_html_url" content="https://www.biorxiv.org/content/10.1101/2024.11.13.623301v1.abstract" /> <meta name="citation_full_html_url" content="https://www.biorxiv.org/content/10.1101/2024.11.13.623301v1.full" /> <meta name="citation_pdf_url" content="https://www.biorxiv.org/content/biorxiv/early/2024/11/15/2024.11.13.623301.full.pdf" /> <meta name="citation_doi" content="10.1101/2024.11.13.623301" /> <meta name="citation_num_pages" content="40" /> <meta name="citation_article_type" content="Article" /> <meta name="citation_section" content="New Results" /> <meta name="citation_firstpage" content="2024.11.13.623301" /> <meta name="citation_author" content="Saima Usman" /> <meta name="citation_author_institution" content="Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London" /> <meta name="citation_author" content="W. Andrew Yeudall" /> <meta name="citation_author_institution" content="Department of Oral Biology and Diagnostic Sciences, The Dental College of Georgia, Augusta University" /> <meta name="citation_author_institution" content="Georgia Cancer Center, Augusta University" /> <meta name="citation_author" content="Muy-Teck Teh" /> <meta name="citation_author_institution" content="Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0002-7725-8355" /> <meta name="citation_author" content="Fatemah Ghloum" /> <meta name="citation_author_institution" content="Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London" /> <meta name="citation_author" content="Hemanth Tummala" /> <meta name="citation_author_institution" content="Centre for Genomics and Child Health, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London" /> <meta name="citation_author" content="Ahmad Waseem" /> <meta name="citation_author_institution" content="Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London" /> <meta name="citation_author_email" content="a.waseem@qmul.ac.uk" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0002-7941-266X" /> <meta name="citation_reference" content="Usman S, Waseem NH, Nguyen TK, Mohsin S, Jamal A, Teh M-T, Waseem A: Vimentin Is at the Heart of Epithelial Mesenchymal Transition (EMT) Mediated Metastasis. 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Oncogene 2023, 42:2074." /> <meta name="twitter:title" content="A single cysteine residue in vimentin regulates long non-coding RNA XIST to suppress epithelial-mesenchymal transition and stemness in breast cancer" /> <meta name="twitter:site" content="@biorxivpreprint" /> <meta name="twitter:card" content="summary" /> <meta name="twitter:image" content="https://www.biorxiv.org/sites/default/files/images/biorxiv_logo_homepage7-5-small.png" /> <meta name="twitter:description" content="Vimentin is a type III intermediate filament (IF) protein, that is induced in a large number of solid tumours. A single cysteine at position 328 in vimentin plays a crucial role in assembly, organisation and stability of IFs. However, its exact function during epithelial mesenchymal transition (EMT) and cancer progression has not been investigated. To investigate this, we have transduced wildtype (WT) and C328S vimentin separately in MCF-7 cells that lack endogenous vimentin. The expression of C328-VIM impacted vimentin-actin interactions and induced EMT-like features that include enhanced cell proliferation, migration, invasion accompanied by reduced cell adhesion when compared to the wildtype cells. Functional transcriptomic studies confirmed the upregulation of EMT and mesenchymal markers, downregulation of epithelial markers as well as acquisition of signatures associated with cancer stemness ( CD56, Oct4, PROCR and CD49f ) thus transforming MCF-7 cells from oestrogen positive to triple reduced ( ESR1, PGR, and HER2 ) status. We also observed a stark increase in the expression of long non-coding RNA, XIST in MCF-7 cells expressing C328-VIM. Targeting the mutant vimentin or XIST by RNA interference partially reversed the phenotypes in C328-VIM expressing MCF-7 cells. Furthermore, introduction of C328-VIM cells into nude mice promoted tumour growth by increasing cancer stemness in an oestrogen independent manner. Altogether, our studies provide insight into how cysteine 328 in vimentin dictates mechano-transduction signals to remodel actin cytoskeleton and protect against EMT and cancer growth via modulating lncRNA XIST . Therefore, targeting vimentin and/or XIST via RNA interference should be a promising therapeutic strategy for breast cancer treatment. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-title" property="og:title" content="A single cysteine residue in vimentin regulates long non-coding RNA XIST to suppress epithelial-mesenchymal transition and stemness in breast cancer" /> <meta name="og-url" property="og:url" content="https://www.biorxiv.org/content/10.1101/2024.11.13.623301v1" /> <meta name="og-site-name" property="og:site_name" content="bioRxiv" /> <meta name="og-description" property="og:description" content="Vimentin is a type III intermediate filament (IF) protein, that is induced in a large number of solid tumours. A single cysteine at position 328 in vimentin plays a crucial role in assembly, organisation and stability of IFs. However, its exact function during epithelial mesenchymal transition (EMT) and cancer progression has not been investigated. To investigate this, we have transduced wildtype (WT) and C328S vimentin separately in MCF-7 cells that lack endogenous vimentin. The expression of C328-VIM impacted vimentin-actin interactions and induced EMT-like features that include enhanced cell proliferation, migration, invasion accompanied by reduced cell adhesion when compared to the wildtype cells. Functional transcriptomic studies confirmed the upregulation of EMT and mesenchymal markers, downregulation of epithelial markers as well as acquisition of signatures associated with cancer stemness ( CD56, Oct4, PROCR and CD49f ) thus transforming MCF-7 cells from oestrogen positive to triple reduced ( ESR1, PGR, and HER2 ) status. We also observed a stark increase in the expression of long non-coding RNA, XIST in MCF-7 cells expressing C328-VIM. Targeting the mutant vimentin or XIST by RNA interference partially reversed the phenotypes in C328-VIM expressing MCF-7 cells. Furthermore, introduction of C328-VIM cells into nude mice promoted tumour growth by increasing cancer stemness in an oestrogen independent manner. Altogether, our studies provide insight into how cysteine 328 in vimentin dictates mechano-transduction signals to remodel actin cytoskeleton and protect against EMT and cancer growth via modulating lncRNA XIST . Therefore, targeting vimentin and/or XIST via RNA interference should be a promising therapeutic strategy for breast cancer treatment. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-type" property="og:type" content="article" /> <meta name="og-image" property="og:image" content="https://www.biorxiv.org/sites/default/files/images/biorxiv_logo_homepage7-5-small.png" /> <meta name="citation_date" content="2024-11-15" /> <link rel="alternate" type="application/vnd.ms-powerpoint" title="Powerpoint" href="/content/10.1101/2024.11.13.623301v1.ppt" /> <meta name="description" content="bioRxiv - the preprint server for biology, operated by Cold Spring Harbor Laboratory, a research and educational institution" /> <meta name="generator" content="Drupal 7 (http://drupal.org)" /> <link rel="canonical" href="https://www.biorxiv.org/content/10.1101/2024.11.13.623301v1" /> <link rel="shortlink" href="https://www.biorxiv.org/node/4226060" /> <title>A 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class="highwire-article-citation highwire-citation-type-highwire-article node4226060" data-node-nid="4226060" id="node-4226060--21163700254" data-pisa="biorxiv;2024.11.13.623301v1" data-pisa-master="biorxiv;2024.11.13.623301" data-apath="/biorxiv/early/2024/11/15/2024.11.13.623301.atom" data-hw-author-tooltip-instance="highwire_author_tooltip"><div class="highwire-cite highwire-cite-highwire-article highwire-citation-biorxiv-article-top clearfix has-author-tooltip" > <span class="biorxiv-article-type"> New Results </span> <h1 class="highwire-cite-title" id="page-title">A single cysteine residue in vimentin regulates long non-coding RNA <em>XIST</em> to suppress epithelial-mesenchymal transition and stemness in breast cancer</h1> <div class="highwire-cite-authors" ><span class="highwire-citation-authors"><span class="highwire-citation-author first" data-delta="0"><span class="nlm-given-names">Saima</span> <span class="nlm-surname">Usman</span></span>, <span class="highwire-citation-author" data-delta="1"><span class="nlm-given-names">W. Andrew</span> <span class="nlm-surname">Yeudall</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="2"><a href="http://orcid.org/0000-0002-7725-8355" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Muy-Teck</span> <span class="nlm-surname">Teh</span></span>, <span class="highwire-citation-author" data-delta="3"><span class="nlm-given-names">Fatemah</span> <span class="nlm-surname">Ghloum</span></span>, <span class="highwire-citation-author" data-delta="4"><span class="nlm-given-names">Hemanth</span> <span class="nlm-surname">Tummala</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="5"><a href="http://orcid.org/0000-0002-7941-266X" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Ahmad</span> <span class="nlm-surname">Waseem</span></span></span></div> <div class="highwire-cite-metadata" ><span class="highwire-cite-metadata-doi highwire-cite-metadata"><span class="label">doi:</span> https://doi.org/10.1101/2024.11.13.623301 </span></div> </div> <div id="hw-article-author-popups-node-4226060--21163700254" style="display: none;"><div class="author-tooltip-0"><div class="author-tooltip-name">Saima Usman </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>1</span><span class='nlm-institution'>Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London</span>, Newark Street, London E1 2AT, <span class='nlm-country'>UK</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Saima%2BUsman%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Usman%20S&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link last"><a href="/search/author1%3ASaima%2BUsman%2B" rel="nofollow" class="" data-icon-position="" data-hide-link-title="0">Search for this author on this site</a></li></ul></div><div class="author-tooltip-1"><div class="author-tooltip-name">W. Andrew Yeudall </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>2</span><span class='nlm-institution'>Department of Oral Biology and Diagnostic Sciences, The Dental College of Georgia, Augusta University</span>, Augusta, GA 30912, <span class='nlm-country'>USA</span></div><div class='author-affiliation'><span class='nlm-sup'>3</span><span class='nlm-institution'>Georgia Cancer Center, Augusta University</span>, Augusta, GA 30912, <span class='nlm-country'>USA</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=W.%2BAndrew%2BYeudall%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Yeudall%20WA&link_type=AUTHORSEARCH" 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class='author-affiliation'><span class='nlm-sup'>1</span><span class='nlm-institution'>Centre for Oral Immunobiology and Regenerative Medicine, Institute of Dentistry, Barts and The London School of Medicine and Dentistry, Queen Mary University of London</span>, Newark Street, London E1 2AT, <span class='nlm-country'>UK</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Fatemah%2BGhloum%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Ghloum%20F&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link last"><a href="/search/author1%3AFatemah%2BGhloum%2B" rel="nofollow" class="" 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href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Ahmad%2BWaseem%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Waseem%20A&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link"><a href="/search/author1%3AAhmad%2BWaseem%2B" rel="nofollow" class="" data-icon-position="" data-hide-link-title="0">Search for this author on this site</a></li><li class="author-orcid-link"><a href="http://orcid.org/0000-0002-7941-266X" target="_blank" class="" data-icon-position="" data-hide-link-title="0">ORCID record for Ahmad Waseem</a></li><li class="author-corresp-email-link last"><span>For correspondence: <a href="mailto:a.waseem@qmul.ac.uk" class="" data-icon-position="" 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solid tumours. A single cysteine at position 328 in vimentin plays a crucial role in assembly, organisation and stability of IFs. However, its exact function during epithelial mesenchymal transition (EMT) and cancer progression has not been investigated. To investigate this, we have transduced wildtype (WT) and C328S vimentin separately in MCF-7 cells that lack endogenous vimentin. The expression of C328-VIM impacted vimentin-actin interactions and induced EMT-like features that include enhanced cell proliferation, migration, invasion accompanied by reduced cell adhesion when compared to the wildtype cells. Functional transcriptomic studies confirmed the upregulation of EMT and mesenchymal markers, downregulation of epithelial markers as well as acquisition of signatures associated with cancer stemness (<em>CD56, Oct4, PROCR and CD49f</em>) thus transforming MCF-7 cells from oestrogen positive to triple reduced (<em>ESR1, PGR, and HER2</em>) status. We also observed a stark increase in the expression of long non-coding RNA, <em>XIST</em> in MCF-7 cells expressing C328-VIM. Targeting the mutant vimentin or <em>XIST</em> by RNA interference partially reversed the phenotypes in C328-VIM expressing MCF-7 cells. Furthermore, introduction of C328-VIM cells into nude mice promoted tumour growth by increasing cancer stemness in an oestrogen independent manner. Altogether, our studies provide insight into how cysteine 328 in vimentin dictates mechano-transduction signals to remodel actin cytoskeleton and protect against EMT and cancer growth via modulating lncRNA <em>XIST</em>. Therefore, targeting vimentin and/or <em>XIST</em> via RNA interference should be a promising therapeutic strategy for breast cancer treatment.</p></div><h3>Competing Interest Statement</h3><p id="p-3">The authors have declared no competing interest.</p><span class="highwire-journal-article-marker-end"></span></div><span class="related-urls"></span></div></div> </div> </div> <div class="panel-separator"></div><div class="panel-pane pane-biorxiv-copyright" > <div class="pane-content"> <div class="field field-name-field-highwire-copyright field-type-text field-label-inline clearfix"><div class="field-label">Copyright </div><div class="field-items"><div class="field-item even">The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.<span class="license-type"> It is made available under a <a href="http://creativecommons.org/licenses/by/4.0/" class="" data-icon-position="" data-hide-link-title="0">CC-BY 4.0 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epithelial-mesenchymal transition and stemness in breast cancer</div> </div> <div class="highwire-cite-authors" ><span class="highwire-citation-authors"><span class="highwire-citation-author first" data-delta="0"><span class="nlm-given-names">Saima</span> <span class="nlm-surname">Usman</span></span>, <span class="highwire-citation-author" data-delta="1"><span class="nlm-given-names">W. 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