MP85-08 DIRECT REGULATION OF COLLAGEN CROSS-LINKING ENZYMES (LOXL2 AND PLOD2) BY TUMOR-SUPPRESSIVE MICRORNA-26A/B IN RENAL CELL CARCINOMA

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology II1 Apr 2016MP85-08 DIRECT REGULATION OF COLLAGEN CROSS-LINKING ENZYMES (LOXL2 AND PLOD2) BY TUMOR-SUPPRESSIVE MICRORNA-26A/B IN RENAL CELL CARCINOMA Akira Kurozumi, Mayuko Kato, Yusuke Goto, Atsushi Okato, Ryosuke Matsushita, Hideki Enokida, Masayuki Nakagawa, Tomohiko Ichikawa, and Naohiko Seki Akira KurozumiAkira Kurozumi More articles by this author , Mayuko KatoMayuko Kato More articles by this author , Yusuke GotoYusuke Goto More articles by this author , Atsushi OkatoAtsushi Okato More articles by this author , Ryosuke MatsushitaRyosuke Matsushita More articles by this author , Hideki EnokidaHideki Enokida More articles by this author , Masayuki NakagawaMasayuki Nakagawa More articles by this author , Tomohiko IchikawaTomohiko Ichikawa More articles by this author , and Naohiko SekiNaohiko Seki More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2274AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Our recent studies of microRNA (miRNA) expression signatures of human cancers revealed that microRNA-26a (miR-26a) and microRNA-26b (miR-26b) were significantly reduced in cancer tissues, suggesting these miRNAs act as tumor-suppressors. The aim of this study was to investigate the functional significance of miR-26a and miR-26b in RCC and to identify miR-26a/b-mediated target genes involved in RCC oncogenesis and metastasis. Our present data showed that collagen cross-linking enzymes were direct regulation of these miRNAs. Collagens are major components of the extracellular matrix (ECM) and aberrant stiffening of ECM enhances cell growth, survival, integrin signaling, and focal adhesion formation. METHODS Expression levels of miR-26a/b and their candidate target genes were evaluated in clear cell RCC (ccRCC) clinical specimens (fifteen radical nephrectomy specimens) and RCC cell lines (786-O and A498) by PCR methods. Gain-of-function studies (cell proliferation, migration and invasion assays) were performed using transfection of mature miR-26a/b into RCC cells. Genome-wide gene expression analysis and in silico analysis were applied to investigate molecular targets regulated by miR-26a/b in RCC cells. A luciferase reporter assay was carried out to determine whether 3′UTR of target genes have actual binding sites for miR-26a/b. RESULTS The expression levels of miR-26a/b were significantly reduced in ccRCC clinical specimens and RCC cell lines compared with non-cancerous kidney tissues (P < 0.05). Restoration of miR-26a/b significantly inhibited cancer cell migration and invasion in RCC cell lines, 786-O and A498 (P < 0.01). Collagen cross-linking enzymes, lysyl oxidase-like 2 (LOXL2) and procollagen Lysyl Hydroxylase 2 (PLOD2) were identified as direct target genes of miR-26a/b by genome-wide gene expression analysis and in silico analysis. Silencing of LOXL2 or PLOD2 significantly reduced cancer cell migration and invasion in RCC cells. CONCLUSIONS Downregulation of miR-26a/b cause to overexpression of collagen cross-linking enzymes and promoting ECM stiffening. Overexpression of LOXL2 and PLOD2 might enhance RCC cell metastasis. Elucidation of tumor-suppressive miR-26a/b regulated molecular pathways and targets could provide new information on potential therapeutic strategies in RCC. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1100 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Akira Kurozumi More articles by this author Mayuko Kato More articles by this author Yusuke Goto More articles by this author Atsushi Okato More articles by this author Ryosuke Matsushita More articles by this author Hideki Enokida More articles by this author Masayuki Nakagawa More articles by this author Tomohiko Ichikawa More articles by this author Naohiko Seki More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...