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1097-41645352014Mar06Molecular cellMol CellThe breast cancer oncogene EMSY represses transcription of antimetastatic microRNA miR-31.806818806-1810.1016/j.molcel.2014.01.029S1097-2765(14)00112-9Amplification of the EMSY gene in sporadic breast and ovarian cancers is a poor prognostic indicator. Although EMSY has been linked to transcriptional silencing, its mechanism of action is unknown. Here, we report that EMSY acts as an oncogene, causing the transformation of cells in vitro and potentiating tumor formation and metastatic features in vivo. We identify an inverse correlation between EMSY amplification and miR-31 expression, an antimetastatic microRNA, in the METABRIC cohort of human breast samples. Re-expression of miR-31 profoundly reduced cell migration, invasion, and colony-formation abilities of cells overexpressing EMSY or haboring EMSY amplification. We show that EMSY is recruited to the miR-31 promoter by the DNA binding factor ETS-1, and it represses miR-31 transcription by delivering the H3K4me3 demethylase JARID1b/PLU-1/KDM5B. Altogether, these results suggest a pathway underlying the role of EMSY in breast cancer and uncover potential diagnostic and therapeutic targets in sporadic breast cancer.Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.ViréEmmanuelleEGurdon Institute and Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.CurtisChristinaCDepartment of Oncology and Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Robinson Way, CB2 0RE, UK.DavalosVeronicaVCancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona 08907, Spain.GitAnnaADepartment of Oncology and Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Robinson Way, CB2 0RE, UK.RobsonSamuelSGurdon Institute and Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.VillanuevaAlbertoACancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona 08907, Spain.VidalAugustADepartment of Pathological Anatomy, Bellvitge University Hospital, Bellvitge Biomedical Research Institute (IDIBELL), Barcelona 08907, Spain.BarbieriIsaiaIAparicioSamuelSDepartment of Molecular Oncology, British Columbia Cancer Agency, 675 West 10(th) Avenue, V5Z 1L3 Vancouver, Canada.EstellerManelMCancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona 08907, Spain.CaldasCarlosCDepartment of Oncology and Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Robinson Way, CB2 0RE, UK; Cambridge Breast Unit, Addenbrooke's Hospital, Cambridge University Hospital, NHS Foundation Trust and NIHR Cambridge Biomedical Research Centre, Cambridge CB2 2QQ, UK; Cambridge Experimental Cancer Medicine Centre (ECMC), Cambridge CB2 0RE, UK.KouzaridesTonyTGurdon Institute and Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK. Electronic address: t.kouzarides@gurdon.cam.ac.uk.eng092096WT_Wellcome TrustUnited Kingdom10827CRUK_Cancer Research UKUnited Kingdom16942CRUK_Cancer Research UKUnited Kingdom17001CRUK_Cancer Research UKUnited KingdomJournal ArticleResearch Support, Non-U.S. Gov't20140227
United StatesMol Cell98025711097-27650EMSY protein, human0MIRN31 microRNA, human0MicroRNAs0Neoplasm Proteins0Nuclear Proteins0RNA, Small Interfering0Repressor ProteinsIMBreast Cancer Res. 2014 Nov 18;16(6):467. doi: 10.1186/s13058-014-0467-x.25927669Mol Cell. 2014 Apr 10;54(1):203. doi: 10.1016/j.molcel.2014.03.041.28898635AnimalsBase SequenceBreast NeoplasmsmetabolismCell MovementCohort StudiesFemaleGene Expression Regulation, NeoplasticGene SilencingHumansMCF-7 CellsMiceMice, NudeMicroRNAsgeneticsphysiologyMolecular Sequence DataNIH 3T3 CellsNeoplasm InvasivenessNeoplasm MetastasisNeoplasm ProteinsmetabolismphysiologyNuclear ProteinsmetabolismphysiologyOncogenesgeneticsRNA, Small InterferingmetabolismRepressor Proteinsmetabolismphysiology20136272013111120141232014346020143460201442960201436ppublish24582497PMC398888610.1016/j.molcel.2014.01.029S1097-2765(14)00112-9Albert M., Schmitz S.U., Kooistra S.M., Malatesta M., Morales Torres C., Rekling J.C., Johansen J.V., Abarrategui I., Helin K. The histone demethylase Jarid1b ensures faithful mouse development by protecting developmental genes from aberrant H3K4me3. PLoS Genet. 2013;9:e1003461.PMC363009323637629Augoff K., McCue B., Plow E.F., Sossey-Alaoui K. miR-31 and its host gene lncRNA LOC554202 are regulated by promoter hypermethylation in triple-negative breast cancer. Mol. Cancer. 2012;11:5.PMC329850322289355Barrett A., Madsen B., Copier J., Lu P.J., Cooper L., Scibetta A.G., Burchell J., Taylor-Papadimitriou J. PLU-1 nuclear protein, which is upregulated in breast cancer, shows restricted expression in normal human adult tissues: a new cancer/testis antigen? Int. J. Cancer. 2002;101:581–588.12237901Barrett A., Santangelo S., Tan K., Catchpole S., Roberts K., Spencer-Dene B., Hall D., Scibetta A., Burchell J., Verdin E. Breast cancer associated transcriptional repressor PLU-1/JARID1B interacts directly with histone deacetylases. Int. J. Cancer. 2007;121:265–275.17373667Brown L.A., Irving J., Parker R., Kim H., Press J.Z., Longacre T.A., Chia S., Magliocco A., Makretsov N., Gilks B. Amplification of EMSY, a novel oncogene on 11q13, in high grade ovarian surface epithelial carcinomas. Gynecol. Oncol. 2006;100:264–270.16236351Brown L.A., Kalloger S.E., Miller M.A., Shih IeM., McKinney S.E., Santos J.L., Swenerton K., Spellman P.T., Gray J., Gilks C.B., Huntsman D.G. Amplification of 11q13 in ovarian carcinoma. Genes Chromosomes Cancer. 2008;47:481–489.18314909Brown L.A., Johnson K., Leung S., Bismar T.A., Benítez J., Foulkes W.D., Huntsman D.G. Co-amplification of CCND1 and EMSY is associated with an adverse outcome in ER-positive tamoxifen-treated breast cancers. Breast Cancer Res. Treat. 2010;121:347–354.19636701Buggy Y., Maguire T.M., McGreal G., McDermott E., Hill A.D., O’Higgins N., Duffy M.J. Overexpression of the Ets-1 transcription factor in human breast cancer. Br. J. Cancer. 2004;91:1308–1315.PMC240990515365563Bullock M.D., Sayan A.E., Packham G.K., Mirnezami A.H. MicroRNAs: critical regulators of epithelial to mesenchymal (EMT) and mesenchymal to epithelial transition (MET) in cancer progression. Biol. Cell. 2012;104:3–12.22188537Catchpole S., Spencer-Dene B., Hall D., Santangelo S., Rosewell I., Guenatri M., Beatson R., Scibetta A.G., Burchell J.M., Taylor-Papadimitriou J. PLU-1/JARID1B/KDM5B is required for embryonic survival and contributes to cell proliferation in the mammary gland and in ER+ breast cancer cells. Int. J. Oncol. 2011;38:1267–1277.21369698Chavali G.B., Ekblad C.M., Basu B.P., Brissett N.C., Veprintsev D., Hughes-Davies L., Kouzarides T., Itzhaki L.S., Doherty A.J. Crystal structure of the ENT domain of human EMSY. J. Mol. Biol. 2005;350:964–973.15978617Costa Y., Ding J., Theunissen T.W., Faiola F., Hore T.A., Shliaha P.V., Fidalgo M., Saunders A., Lawrence M., Dietmann S. NANOG-dependent function of TET1 and TET2 in establishment of pluripotency. Nature. 2013;495:370–374.PMC360664523395962Curtis C., Shah S.P., Chin S.F., Turashvili G., Rueda O.M., Dunning M.J., Speed D., Lynch A.G., Samarajiwa S., Yuan Y., METABRIC Group The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature. 2012;486:346–352.PMC344084622522925Dawson M.A., Bannister A.J., Göttgens B., Foster S.D., Bartke T., Green A.R., Kouzarides T. JAK2 phosphorylates histone H3Y41 and excludes HP1alpha from chromatin. Nature. 2009;461:819–822.PMC378514719783980Dvinge H., Git A., Gräf S., Salmon-Divon M., Curtis C., Sottoriva A., Zhao Y., Hirst M., Armisen J., Miska E.A. The shaping and functional consequences of the microRNA landscape in breast cancer. Nature. 2013;497:378–382.23644459Ekblad C.M., Chavali G.B., Basu B.P., Freund S.M., Veprintsev D., Hughes-Davies L., Kouzarides T., Doherty A.J., Itzhaki L.S. Binding of EMSY to HP1beta: implications for recruitment of HP1beta and BS69. EMBO Rep. 2005;6:675–680.PMC136910715947784Ezell S.A., Polytarchou C., Hatziapostolou M., Guo A., Sanidas I., Bihani T., Comb M.J., Sourvinos G., Tsichlis P.N. The protein kinase Akt1 regulates the interferon response through phosphorylation of the transcriptional repressor EMSY. Proc. Natl. Acad. Sci. USA. 2012;109:E613–E621.PMC330977322315412Furlan A., Vercamer C., Desbiens X., Pourtier A. Ets-1 triggers and orchestrates the malignant phenotype of mammary cancer cells within their matrix environment. J. Cell. Physiol. 2008;215:782–793.18181172Garapaty S., Xu C.F., Trojer P., Mahajan M.A., Neubert T.A., Samuels H.H. Identification and characterization of a novel nuclear protein complex involved in nuclear hormone receptor-mediated gene regulation. J. Biol. Chem. 2009;284:7542–7552.PMC265804919131338Garzon R., Fabbri M., Cimmino A., Calin G.A., Croce C.M. MicroRNA expression and function in cancer. Trends Mol. Med. 2006;12:580–587.17071139Garzon R., Calin G.A., Croce C.M. MicroRNAs in Cancer. Annu. Rev. Med. 2009;60:167–179.19630570Gilles C., Polette M., Birembaut P., Brünner N., Thompson E.W. Expression of c-ets-1 mRNA is associated with an invasive, EMT-derived phenotype in breast carcinoma cell lines. Clin. Exp. Metastasis. 1997;15:519–526.9247254Hughes-Davies L., Huntsman D., Ruas M., Fuks F., Bye J., Chin S.F., Milner J., Brown L.A., Hsu F., Gilks B. EMSY links the BRCA2 pathway to sporadic breast and ovarian cancer. Cell. 2003;115:523–535.14651845Iorio M.V., Croce C.M. MicroRNAs in cancer: small molecules with a huge impact. J. Clin. Oncol. 2009;27:5848–5856.PMC279300319884536Iorio M.V., Ferracin M., Liu C.G., Veronese A., Spizzo R., Sabbioni S., Magri E., Pedriali M., Fabbri M., Campiglio M. MicroRNA gene expression deregulation in human breast cancer. Cancer Res. 2005;65:7065–7070.16103053Kozaki K., Imoto I., Mogi S., Omura K., Inazawa J. Exploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in oral cancer. Cancer Res. 2008;68:2094–2105.18381414Lu P.J., Sundquist K., Baeckstrom D., Poulsom R., Hanby A., Meier-Ewert S., Jones T., Mitchell M., Pitha-Rowe P., Freemont P., Taylor-Papadimitriou J. A novel gene (PLU-1) containing highly conserved putative DNA/chromatin binding motifs is specifically up-regulated in breast cancer. J. Biol. Chem. 1999;274:15633–15645.10336460Lujambio A., Calin G.A., Villanueva A., Ropero S., Sánchez-Céspedes M., Blanco D., Montuenga L.M., Rossi S., Nicoloso M.S., Faller W.J. A microRNA DNA methylation signature for human cancer metastasis. Proc. Natl. Acad. Sci. USA. 2008;105:13556–13561.PMC252887218768788Malovannaya A., Lanz R.B., Jung S.Y., Bulynko Y., Le N.T., Chan D.W., Ding C., Shi Y., Yucer N., Krenciute G. Analysis of the human endogenous coregulator complexome. Cell. 2011;145:787–799.PMC313108321620140Mirnezami A.H., Pickard K., Zhang L., Primrose J.N., Packham G. MicroRNAs: key players in carcinogenesis and novel therapeutic targets. Eur. J. Surg. Oncol. 2009;35:339–347.18644693Moshkin Y.M., Kan T.W., Goodfellow H., Bezstarosti K., Maeda R.K., Pilyugin M., Karch F., Bray S.J., Demmers J.A., Verrijzer C.P. Histone chaperones ASF1 and NAP1 differentially modulate removal of active histone marks by LID-RPD3 complexes during NOTCH silencing. Mol. Cell. 2009;35:782–793.19782028Raouf A., Brown L., Vrcelj N., To K., Kwok W., Huntsman D., Eaves C.J. Genomic instability of human mammary epithelial cells overexpressing a truncated form of EMSY. J. Natl. Cancer Inst. 2005;97:1302–1306.16145051Rodriguez C., Hughes-Davies L., Vallès H., Orsetti B., Cuny M., Ursule L., Kouzarides T., Theillet C. Amplification of the BRCA2 pathway gene EMSY in sporadic breast cancer is related to negative outcome. Clin. Cancer Res. 2004;10:5785–5791.15355907Saito Y., Liang G., Egger G., Friedman J.M., Chuang J.C., Coetzee G.A., Jones P.A. Specific activation of microRNA-127 with downregulation of the proto-oncogene BCL6 by chromatin-modifying drugs in human cancer cells. Cancer Cell. 2006;9:435–443.16766263Switzer C.H., Cheng R.Y., Ridnour L.A., Glynn S.A., Ambs S., Wink D.A. Ets-1 is a transcriptional mediator of oncogenic nitric oxide signaling in estrogen receptor-negative breast cancer. Breast Cancer Res. 2012;14:R125.PMC405310222971289Valastyan S., Weinberg R.A. miR-31: a crucial overseer of tumor metastasis and other emerging roles. Cell Cycle. 2010;9:2124–2129.20505365Valastyan S., Weinberg R.A. Roles for microRNAs in the regulation of cell adhesion molecules. J. Cell Sci. 2011;124:999–1006.PMC305660221402873Valastyan S., Benaich N., Chang A., Reinhardt F., Weinberg R.A. Concomitant suppression of three target genes can explain the impact of a microRNA on metastasis. Genes Dev. 2009;23:2592–2597.PMC277976319875476Valastyan S., Reinhardt F., Benaich N., Calogrias D., Szász A.M., Wang Z.C., Brock J.E., Richardson A.L., Weinberg R.A. A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell. 2009;137:1032–1046.PMC276660919524507Valastyan S., Chang A., Benaich N., Reinhardt F., Weinberg R.A. Concurrent suppression of integrin alpha5, radixin, and RhoA phenocopies the effects of miR-31 on metastasis. Cancer Res. 2010;70:5147–5154.PMC289135020530680Valastyan S., Chang A., Benaich N., Reinhardt F., Weinberg R.A. Activation of miR-31 function in already-established metastases elicits metastatic regression. Genes Dev. 2011;25:646–659.PMC305983721406558Visone R., Croce C.M. MiRNAs and cancer. Am. J. Pathol. 2009;174:1131–1138.PMC267134619264914Vrba L., Muñoz-Rodríguez J.L., Stampfer M.R., Futscher B.W. miRNA gene promoters are frequent targets of aberrant DNA methylation in human breast cancer. PLoS ONE. 2013;8:e54398.PMC354703323342147Yamagishi M., Nakano K., Miyake A., Yamochi T., Kagami Y., Tsutsumi A., Matsuda Y., Sato-Otsubo A., Muto S., Utsunomiya A. Polycomb-mediated loss of miR-31 activates NIK-dependent NF-κB pathway in adult T cell leukemia and other cancers. Cancer Cell. 2012;21:121–135.22264793Yamane K., Tateishi K., Klose R.J., Fang J., Fabrizio L.A., Erdjument-Bromage H., Taylor-Papadimitriou J., Tempst P., Zhang Y. PLU-1 is an H3K4 demethylase involved in transcriptional repression and breast cancer cell proliferation. Mol. Cell. 2007;25:801–812.17363312Yao J., Polyak K. EMSY links breast cancer gene 2 to the ‘Royal Family’. Breast Cancer Res. 2004;6:201–203.PMC54917715318925