{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,14]],"date-time":"2026-06-14T08:48:54Z","timestamp":1781426934344,"version":"3.54.1"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:00:00Z","timestamp":1760486400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["R01CA267872"],"award-info":[{"award-number":["R01CA267872"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100008258","name":"Hyundai Hope On Wheels","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100008258","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>The STAG2 tumor suppressor gene is commonly inactivated by mutations in a wide range of common cancer types. STAG2 encodes a component of the cohesin complex, which controls sister chromatid cohesion and 3D genome organization. In bladder cancer, STAG2 mutations are most common in the earliest low-grade lesions, suggesting that mutational inactivation of STAG2 may be an initiating event. To provide insight into the mechanisms of STAG2 tumor suppression in bladder cancer, siRNA and shRNA were used to knock down STAG2 in several different human non-neoplastic bladder cancer precursor cell lines. Gene editing was used to generate cultured human cancer cell lines that differ only in the presence or absence of bladder-cancer derived STAG2 mutations. These systems were interrogated using RNA-seq, Western blot, and qRT-PCR before and after induced differentiation. We find that inactivation of STAG2 in bladder cancer cells and in bladder epithelial precursor cells resulted in concomitant inactivation of the H3K27me3 Polycomb chromatin mark. Inactivation of STAG2 also attenuated induced differentiation of bladder epithelial precursor cells. STAG2 and other components of cohesin were upregulated during this differentiation process. This study provides new insights into the role of STAG2 in the pathogenesis of bladder cancer, demonstrating roles for STAG2 in the regulation of Polycomb-mediated epigenetic regulation and in the differentiation of bladder epithelial precursor cells.<\/jats:p>","DOI":"10.1371\/journal.pone.0333128","type":"journal-article","created":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T17:24:58Z","timestamp":1760549098000},"page":"e0333128","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["STAG2 regulates polycomb and differentiation in urothelial precursors and bladder cancer"],"prefix":"10.1371","volume":"20","author":[{"given":"Youngrok","family":"Park","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanying","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0381-5833","authenticated-orcid":true,"given":"Tianyi","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Hill","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyuan","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Sadzewicz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luke","family":"Tallon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung-Sik","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fulai","family":"Jin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8126-7764","authenticated-orcid":true,"given":"Todd","family":"Waldman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,10,15]]},"reference":[{"key":"pone.0333128.ref001","first-page":"10","article-title":"Cancer statistics, 2005","volume":"55","author":"A Jemal","year":"2005","journal-title":"CA Cancer J Clin"},{"issue":"2","key":"pone.0333128.ref002","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.ccr.2004.08.002","article-title":"Focus on bladder cancer","volume":"6","author":"CPN Dinney","year":"2004","journal-title":"Cancer Cell"},{"key":"pone.0333128.ref003","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1146\/annurev-pathol-012513-104703","article-title":"Origins of Bladder Cancer","volume":"11","author":"B Czerniak","year":"2016","journal-title":"Annu Rev Pathol"},{"issue":"11","key":"pone.0333128.ref004","article-title":"Sister chromatid cohesion","volume":"4","author":"J-M Peters","year":"2012","journal-title":"Cold Spring Harb Perspect Biol"},{"issue":"6045","key":"pone.0333128.ref005","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1126\/science.1203619","article-title":"Mutational inactivation of STAG2 causes aneuploidy in human cancer","volume":"333","author":"DA Solomon","year":"2011","journal-title":"Science"},{"issue":"1","key":"pone.0333128.ref006","first-page":"1","article-title":"Cohesin mutations in human cancer","volume":"1866","author":"VK Hill","year":"2016","journal-title":"Biochim Biophys Acta"},{"issue":"9","key":"pone.0333128.ref007","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1038\/s41568-020-0270-1","article-title":"Emerging themes in cohesin cancer biology","volume":"20","author":"T Waldman","year":"2020","journal-title":"Nat Rev Cancer"},{"issue":"12","key":"pone.0333128.ref008","doi-asserted-by":"crossref","first-page":"1428","DOI":"10.1038\/ng.2800","article-title":"Frequent truncating mutations of STAG2 in bladder cancer","volume":"45","author":"DA Solomon","year":"2013","journal-title":"Nat Genet"},{"issue":"12","key":"pone.0333128.ref009","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1038\/ng.2799","article-title":"Recurrent inactivation of STAG2 in bladder cancer is not associated with aneuploidy","volume":"45","author":"C Balb\u00e1s-Mart\u00ednez","year":"2013","journal-title":"Nat Genet"},{"issue":"12","key":"pone.0333128.ref010","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.1038\/ng.2798","article-title":"Whole-genome and whole-exome sequencing of bladder cancer identifies frequent alterations in genes involved in sister chromatid cohesion and segregation","volume":"45","author":"G Guo","year":"2013","journal-title":"Nat Genet"},{"issue":"17","key":"pone.0333128.ref011","doi-asserted-by":"crossref","first-page":"4145","DOI":"10.1158\/1078-0432.CCR-17-3244","article-title":"STAG2 is a biomarker for prediction of recurrence and progression in papillary non-muscle-invasive bladder cancer","volume":"24","author":"A Lelo","year":"2018","journal-title":"Clin Cancer Res"},{"issue":"7","key":"pone.0333128.ref012","doi-asserted-by":"crossref","DOI":"10.1016\/j.urolonc.2021.02.007","article-title":"STAG2 as a prognostic biomarker in low-grade non-muscle invasive bladder cancer","volume":"39","author":"A Taber","year":"2021","journal-title":"Urol Oncol"},{"issue":"3","key":"pone.0333128.ref013","article-title":"Cohesin disrupts polycomb-dependent chromosome interactions in embryonic stem cells","volume":"30","author":"JDP Rhodes","year":"2020","journal-title":"Cell Rep"},{"issue":"22","key":"pone.0333128.ref014","doi-asserted-by":"crossref","first-page":"4172","DOI":"10.1242\/dev.084566","article-title":"Wapl antagonizes cohesin binding and promotes Polycomb-group silencing in Drosophila","volume":"139","author":"MD Cunningham","year":"2012","journal-title":"Development"},{"issue":"6","key":"pone.0333128.ref015","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pgen.1003560","article-title":"Cohesin and polycomb proteins functionally interact to control transcription at silenced and active genes","volume":"9","author":"CA Schaaf","year":"2013","journal-title":"PLoS Genet"},{"issue":"4","key":"pone.0333128.ref016","article-title":"Polycomb group proteins regulate chromatin architecture in mouse oocytes and early embryos","volume":"77","author":"Z Du","year":"2020","journal-title":"Mol Cell"},{"issue":"6","key":"pone.0333128.ref017","doi-asserted-by":"crossref","first-page":"107341","DOI":"10.1016\/j.jbc.2024.107341","article-title":"STAG2 mutations regulate 3D genome organization, chromatin loops, and Polycomb signaling in glioblastoma multiforme","volume":"300","author":"W Xu","year":"2024","journal-title":"J Biol Chem"},{"issue":"3","key":"pone.0333128.ref018","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1159\/000087804","article-title":"Increased expression of EZH2, a polycomb group protein, in bladder carcinoma","volume":"75","author":"S Arisan","year":"2005","journal-title":"Urol Int"},{"issue":"5","key":"pone.0333128.ref019","doi-asserted-by":"crossref","first-page":"464","DOI":"10.4161\/epi.24532","article-title":"Multifaceted role of EZH2 in breast and prostate tumorigenesis: epigenetics and beyond","volume":"8","author":"G Deb","year":"2013","journal-title":"Epigenetics"},{"issue":"6","key":"pone.0333128.ref020","doi-asserted-by":"crossref","DOI":"10.1016\/j.ccell.2021.05.007","article-title":"STAG2 loss rewires oncogenic and developmental programs to promote metastasis in Ewing sarcoma","volume":"39","author":"B Adane","year":"2021","journal-title":"Cancer Cell"},{"issue":"11","key":"pone.0333128.ref021","doi-asserted-by":"crossref","first-page":"1833","DOI":"10.1084\/jem.20151323","article-title":"Cohesin loss alters adult hematopoietic stem cell homeostasis, leading to myeloproliferative neoplasms","volume":"212","author":"J Mullenders","year":"2015","journal-title":"J Exp Med"},{"issue":"6","key":"pone.0333128.ref022","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1016\/j.stem.2015.09.017","article-title":"Leukemia-associated cohesin mutants dominantly enforce stem cell programs and impair human hematopoietic progenitor differentiation","volume":"17","author":"C Mazumdar","year":"2015","journal-title":"Cell Stem Cell"},{"issue":"11","key":"pone.0333128.ref023","doi-asserted-by":"crossref","first-page":"1819","DOI":"10.1084\/jem.20151317","article-title":"Dose-dependent role of the cohesin complex in normal and malignant hematopoiesis","volume":"212","author":"AD Viny","year":"2015","journal-title":"J Exp Med"},{"issue":"12","key":"pone.0333128.ref024","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1016\/j.celrep.2016.02.082","article-title":"Genome-wide RNAi screen identifies cohesin genes as modifiers of renewal and differentiation in human HSCs","volume":"14","author":"R Galeev","year":"2016","journal-title":"Cell Rep"},{"issue":"4","key":"pone.0333128.ref025","doi-asserted-by":"crossref","DOI":"10.1016\/j.stem.2017.07.015","article-title":"Multiplex CRISPR\/Cas9-based genome editing in human hematopoietic stem cells models clonal hematopoiesis and myeloid neoplasia","volume":"21","author":"Z Tothova","year":"2017","journal-title":"Cell Stem Cell"},{"issue":"9","key":"pone.0333128.ref026","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1038\/s41590-018-0184-1","article-title":"Control of inducible gene expression links cohesin to hematopoietic progenitor self-renewal and differentiation","volume":"19","author":"S Cuartero","year":"2018","journal-title":"Nat Immunol"},{"issue":"19","key":"pone.0333128.ref027","doi-asserted-by":"crossref","first-page":"11005","DOI":"10.1093\/nar\/gkab864","article-title":"STAG2 loss-of-function affects short-range genomic contacts and modulates the basal-luminal transcriptional program of bladder cancer cells","volume":"49","author":"L Richart","year":"2021","journal-title":"Nucleic Acids Res"},{"issue":"4","key":"pone.0333128.ref028","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1038\/nmeth.3317","article-title":"HISAT: a fast spliced aligner with low memory requirements","volume":"12","author":"D Kim","year":"2015","journal-title":"Nat Methods"},{"issue":"7","key":"pone.0333128.ref029","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1093\/bioinformatics\/btt656","article-title":"featureCounts: an efficient general purpose program for assigning sequence reads to genomic features","volume":"30","author":"Y Liao","year":"2014","journal-title":"Bioinformatics"},{"issue":"12","key":"pone.0333128.ref030","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"MI Love","year":"2014","journal-title":"Genome Biol"},{"issue":"19","key":"pone.0333128.ref031","article-title":"Epitope tagging of endogenous genes in diverse human cell lines","volume":"36","author":"J-S Kim","year":"2008","journal-title":"Nucleic Acids Res"},{"issue":"2","key":"pone.0333128.ref032","article-title":"Intact cohesion, anaphase, and chromosome segregation in human cells harboring tumor-derived mutations in STAG2","volume":"12","author":"J-S Kim","year":"2016","journal-title":"PLoS Genet"},{"issue":"43","key":"pone.0333128.ref033","doi-asserted-by":"crossref","first-page":"15545","DOI":"10.1073\/pnas.0506580102","article-title":"Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles","volume":"102","author":"A Subramanian","year":"2005","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"7200","key":"pone.0333128.ref034","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/nature07056","article-title":"Dissecting direct reprogramming through integrative genomic analysis","volume":"454","author":"TS Mikkelsen","year":"2008","journal-title":"Nature"},{"issue":"4","key":"pone.0333128.ref035","doi-asserted-by":"crossref","first-page":"449","DOI":"10.3233\/BLC-160065","article-title":"The new immortalized uroepithelial cell line HBLAK contains defined genetic aberrations typical of early stage urothelial tumors","volume":"2","author":"MJ Hoffmann","year":"2016","journal-title":"Bladder Cancer"},{"issue":"4","key":"pone.0333128.ref036","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1101\/gr.253211.119","article-title":"Redundant and specific roles of cohesin STAG subunits in chromatin looping and transcriptional control","volume":"30","author":"V Casa","year":"2020","journal-title":"Genome Res"},{"issue":"24","key":"pone.0333128.ref037","doi-asserted-by":"crossref","first-page":"3573","DOI":"10.15252\/embj.201798004","article-title":"Topologically associating domains and chromatin loops depend on cohesin and are regulated by CTCF, WAPL, and PDS5 proteins","volume":"36","author":"G Wutz","year":"2017","journal-title":"EMBO J"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0333128","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T17:25:04Z","timestamp":1760549104000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0333128"}},"subtitle":[],"editor":[{"given":"Alexis G. Murillo","family":"Carrasco","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2025,10,15]]},"references-count":37,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,10,15]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0333128","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,15]]}}}