You have accessJournal of UrologyBladder Cancer: Basic Research (II)1 Apr 2013927 MOLECULAR ROLE OF MIR-195/497 CLUSTER IN HUMAN BLADDER CANCER BASED ON MULTIPLE DATASET OF MICRORNA EXPRESSION SIGNATURES BY DEEP SEQUENCING Toshihiko Itesako, Hideki Enokida, Tomokazu Yonezawa, Takeshi Yamasaki, Hideo Hidaka, Hirofumi Yoshino, Naohiko Seki, and Masayuki Nakagawa Toshihiko ItesakoToshihiko Itesako Kagoshima, Japan More articles by this author , Hideki EnokidaHideki Enokida Kagoshima, Japan More articles by this author , Tomokazu YonezawaTomokazu Yonezawa Kagoshima, Japan More articles by this author , Takeshi YamasakiTakeshi Yamasaki Kagoshima, Japan More articles by this author , Hideo HidakaHideo Hidaka Kagoshima, Japan More articles by this author , Hirofumi YoshinoHirofumi Yoshino Kagoshima, Japan More articles by this author , Naohiko SekiNaohiko Seki Chiba, Japan More articles by this author , and Masayuki NakagawaMasayuki Nakagawa Kagoshima, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.504AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Recent our miRNA expression signature of human bladder cancer (BC) by deep sequencing revealed new candidate microRNAs (miRNAs) as tumor suppressors. However, a single miRNA signature from a single study might be unreliable for discriminating its significance. In this study, we adopted in-silico analyses using multiple dataset of miRNAs and mRNA gene expression signatures from the published data and our data to determine tumor suppressive miRNAs and their target genes/molecular pathways in BC. METHODS We established a miRNA expression signature in 5 BCs and 5 normal bladder epitheliums (NBEs). We also reviewed the miRNAs expression in other deep sequencing studies, which containing a total of 48 BCs and 38 NBEs. To identify the pathways potentially regulated by the miRNAs, we applied TargetScan database and GENECODIS software, which assigned many putative miRNA targets to known pathways in KEGG [http://www.genome.jp/kegg/pathway.html]. Gene expression analyses of all candidate genes were analyzed by using Gene Expression Omnibus (GEO) database. In these microarray expression data, we examined 90 BCs and 6 NBEs. We performed gain-of-function studies (XTT, wound healing, and cell invasion assay) by the mature miRNAs transfection into BC cell lines. RESULTS Multi-dataset of miRNA deep sequencing analysis revealed that 53 miRNAs were significantly down-regulated in BC. Among the miRNA clusters in the miRNAs list, we focused on miR-195/497 cluster because it has not been fully investigated in BC. The expression of miR-195 and miR-497 were significantly reduced in BCs. Significant inhibitions of cell proliferation, migration, and invasion were observed in the miR-195 and miR-497 transfectants (each, P<0.001). TargetScan algorithm exhibited 6730 putative genes targeted by miR-195 and miR-497. GENECODIS software assigned these genes into 113 significantly enriched signaling pathways. In the top 21 enriched pathways, 98 genes were actually up-regulated in BC according to the expression data of GEO. In particular, RAF1, MEK1, GRB2 and PIK3R2, which are the key molecule in oncogenic pathways, were highly expressed across more than 12 enriched pathways. CONCLUSIONS Our results suggested that oncogenic RAF1, MEK1, GRB2, and PIK3R2 might be significant target genes of tumor suppressive miR-195/497 cluster and that activated RAF-MEK-ERK pathway and PI3K pathway might be attribute to the down regulation of miR-195/497 cluster in BC. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e381 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Toshihiko Itesako Kagoshima, Japan More articles by this author Hideki Enokida Kagoshima, Japan More articles by this author Tomokazu Yonezawa Kagoshima, Japan More articles by this author Takeshi Yamasaki Kagoshima, Japan More articles by this author Hideo Hidaka Kagoshima, Japan More articles by this author Hirofumi Yoshino Kagoshima, Japan More articles by this author Naohiko Seki Chiba, Japan More articles by this author Masayuki Nakagawa Kagoshima, Japan More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...