463 TUMOR SUPPRESSIVE MICRORNA-138 CONTRIBUTES TO INHIBITION OF BOTH CELL MIGRATION AND INVASION THROUGH TARGETING VIMENTIN IN RENAL CELL CARCINOMA

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research (III)1 Apr 2013463 TUMOR SUPPRESSIVE MICRORNA-138 CONTRIBUTES TO INHIBITION OF BOTH CELL MIGRATION AND INVASION THROUGH TARGETING VIMENTIN IN RENAL CELL CARCINOMA Takeshi Yamasaki, Naohiko Seki, Hirofumi Yoshino, Hideo Hidaka, Hideki Enokida, and Masayuki Nakagawa Takeshi YamasakiTakeshi Yamasaki Kagoshima, Japan More articles by this author , Naohiko SekiNaohiko Seki Chiba, Japan More articles by this author , Hirofumi YoshinoHirofumi Yoshino Kagoshima, Japan More articles by this author , Hideo HidakaHideo Hidaka Kagoshima, Japan More articles by this author , Hideki EnokidaHideki Enokida Kagoshima, 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.2013.02.1854AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Our recent miRNA expression signature of renal cell carcinoma (RCC) revealed that microRNA-138 (miR-138) expression was significantly reduced in RCC, suggesting that miR-138 was a candidate tumor suppressor. Recent publications showed that miR-138 might regulate genes related to epithelial-mesenchymal transition (EMT) in several cancers. The aim of study is to investigate the functional significance of miR-138 and to identify its target genes in RCC. METHODS We evaluated miR-138 expression in 33 RCC clinical specimens and adjacent normal kidney tissues by stem-loop RT-PCR. We observed morphologic changes in the miR-138-transfected RCC cell lines (A498 and 786-O). We performed gain-of-function studies such as cell migration and cell invasion assays by using miR-138 transfectants (TFs). Gene expression analysis of the miR-138 TFs and RCC clinical specimens by oligo-microarray was carried out to identify molecular targets of miR-138. TargetScan database implied that vimentin (VIM), an EMT-related gene, was a promising candidate target gene of miR-138. We investigated whether VIM was regulated by miR-138 by real-time PCR and Western blotting. Loss-of-function studies using si-VIM were performed to investigate the functional significance of VIM. We evaluated the expression levels of both VIM mRNA and protein by qRT-PCR and immunohistochemistry. RESULTS The expression levels of miR-138 were significantly reduced in the RCC specimens compared with the normal tissues. Restoration of mature miR-138 in A498 and 786-O caused changes in the bleb-like cell morphology, as characteristics of EMT. Significant inhibitions of cell migration and cell invasion were observed in the miR-138 TFs. The expression levels of VIM expression was suppressed in the miR-138 TFs at both mRNA and protein levels. Loss-of-function studies demonstrated that significant inhibition of cell migration and invasion occurred in the si-VIM transfectants. In clinical RCC specimens, the expression level of VIM was significantly upregulated in comparison with adjacent normal tissues. Furthermore, immunohistochemistry showed that VIM expression levels in RCC specimens were significantly higher than those in the normal tissues. CONCLUSIONS These data suggest that VIM might function as an oncogene and might be regulated by tumor suppressive miR-138. Tumor suppressive miR-138-mediated EMT pathway provides new insights into the potential mechanisms of RCC oncogenesis and metastasis. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e189-e190 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Takeshi Yamasaki Kagoshima, Japan More articles by this author Naohiko Seki Chiba, Japan More articles by this author Hirofumi Yoshino Kagoshima, Japan More articles by this author Hideo Hidaka Kagoshima, Japan More articles by this author Hideki Enokida Kagoshima, Japan More articles by this author Masayuki Nakagawa Kagoshima, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...