You have accessJournal of UrologyBladder Cancer: Basic Research I1 Apr 2014MP21-20 DEVELOPMENT OF THE ORTHOTROPIC MOUSE MODEL OF BLADDER CANCER METASTASIS: THE FUNCTIONAL SIGNIFICANCE OF MICRORNA-218 Shuichi Tatarano, Takeshi Chiyomaru, Tomoaki Ishihara, Satoru Inoguchi, Hideki Enokida, Naohiko Seki, and Masayuki Nakagawa Shuichi TataranoShuichi Tatarano More articles by this author , Takeshi ChiyomaruTakeshi Chiyomaru More articles by this author , Tomoaki IshiharaTomoaki Ishihara More articles by this author , Satoru InoguchiSatoru Inoguchi More articles by this author , Hideki EnokidaHideki Enokida More articles by this author , Naohiko SekiNaohiko Seki More articles by this author , and Masayuki NakagawaMasayuki Nakagawa More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.848AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES We have developed an orthotropic mouse model of bladder cancer (BC) metastasis for evaluating antitumor effects by tumor suppressive microRNAs. We previously demonstrated that miR-218 was a tumor suppressive miRNA and was frequently down-regulated in clinical BC specimens. In vitro examinations demonstrated that the miR-218 transfection brought strong antitumor effects by inhibiting cell migration and invasion as well as inducing cell apoptosis. The aim of this study is to investigate antitumor effects of miR-218 in vivo. METHODS We transfected green fluorescent protein (GFP) to a BC cell line, BOY that had been previously established from metastatic BC spread to lung. BOY cells that were permanently transfected with miR-218 or miR-control were subcutaneously injected into female donor mice (BALB/c nu/nu). Upon successful establishment of tumors (∼500 mm3), tumors were harvested and cut into smaller fragments (∼2 mm/40 mg) for surgical orthotopic implantation into bladder surface of recipient mice for in vivo imaging study. A total of 22 nude mice (11-miR-218, 11-miR-control) were used and tumor growth was examined over the course of 42 days. Emitted fluorescence from GFP was captured on the computer and quantified once a week after implantation. Each three mice were sacrificed to obtain orthotropic and metastatic specimens at the day 42. The remains were kept until they died to evaluate overall survival. RESULTS The fluorescent signal intensity of orthotropic graft and metastatic region was significantly lower in the mice with miR-218-transfected BOY graft compared to the mice with miR-control-transfectant after the day 14 from implantation (77.6 vs. 265.4/mm2 at the day 42, P < 0.0001). There was a trend to significance about the signal intensity of metastatic region including liver and lung between the groups (1.0 vs. 6.6/mm2, P = 0.0547). At the day 56, all the mice with miR-control-transfectant were died, and Kaplan-Meier analysis revealed significant prolonged survival was observed in the mice with miR-218 transfectant (P <0.001). Gene expression analyses of the orthotropic grafts by microarray revealed that several epithelial mesenchymal transition (EMT)-related genes were markedly down regulated in the mice with miR-218 transfectant. CONCLUSIONS There was significant inhibition of in vivo tumor growth and metastasis through miR-218 transfection. Because transfection of miR-218 contributed prolonged survival, this microRNA might be a promising candidate for nucleic acid therapy for the patient with advanced and metastatic BC. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e233 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Shuichi Tatarano More articles by this author Takeshi Chiyomaru More articles by this author Tomoaki Ishihara More articles by this author Satoru Inoguchi More articles by this author Hideki Enokida More articles by this author Naohiko Seki More articles by this author Masayuki Nakagawa More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...