title: MicroRNA-185 inhibits cell proliferation and induces cell apoptosis by targeting VEGFA directly in von Hippel-Lindau-inactivated clear cell renal cell carcinoma.
abstract: OBJECTIVE
  The von Hippel-Lindau (VHL) gene acts as a tumor suppressor in most clear cell renal cell carcinomas (ccRCCs). Tumor growth in ccRCCs relies on many factors that result from the loss of VHL. This study aims to identify new microRNAs with therapeutic potential for VHL-inactivated ccRCCs.
  
  
  METHODS
  We used 786O, A498 (VHL inactivated), and Caki-1 (VHL intact) ccRCC cell lines and 40 ccRCC samples and their adjacent nontumor tissues to measure the expression of microRNA-185 (miR-185) by real-time quantitative polymerase chain reaction. Overexpression or knockdown of VEGFA expression in renal cancer cells was fulfilled by transfecting expression plasmids or small interfering RNAs. Overexpression of miR-185 in ccRCC cell lines was fulfilled by transfecting chemically synthesized miR-185 mimics. The effects of miR-185 on ccRCC cell lines were detected by MTS assay, colony formation assay, and flow cytometric analysis.
  
  
  RESULTS
  Compared with adjacent nontumor renal tissues, miR-185 expression levels decreased significantly in ccRCC tissues. The expression of miR-185 had a negative correlation with tumor size, Fuhrman grade, and TNM staging. Luciferase assay showed that VEGFA was a direct target gene of miR-185. The overexpression of miR-185 significantly inhibited cell proliferation and induced cell apoptosis by down-regulating VEGFA expression in VHL-inactivated ccRCC cells.
  
  
  CONCLUSIONS
  Our results suggest that the miR-185, as a tumor suppressor, plays a pivotal role by inhibiting VEGFA in VHL-inactivated ccRCC.
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Donglai
authors.initials: D
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Hongzhao
authors.initials: H
authors.lastname: Li
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Lv
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Qingbo
authors.initials: Q
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Xintao
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Liangyou
authors.initials: L
authors.lastname: Gu
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China.
authors.forename: Shaoxi
authors.initials: S
authors.lastname: Xiu
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China; Medical School, Nankai University, Tianjin, P.R. China.
authors.forename: Xu
authors.initials: X
authors.lastname: Bao
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China; Medical School, Nankai University, Tianjin, P.R. China.
authors.forename: Junyao
authors.initials: J
authors.lastname: Duan
authors.affiliation: State Key Laboratory of Kidney Diseases, Department of Urology, Chinese People's Liberation Army Medical School, Chinese People's Liberation Army General Hospital, Beijing, P.R. China. Electronic address: xzhang@foxmail.com.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000368:Aged; D017209:Apoptosis / Q000235:genetics; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000473:pathology; D049109:Cell Proliferation / Q000235:genetics; D015536:Down-Regulation; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D055785:Gene Knockdown Techniques; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics* / Q000473:pathology; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D034741:RNA, Small Interfering; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis*; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics
chemical_list: C552431:MIRN185 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human
keywords: Apoptosis; Clear cell renal cell carcinoma; MicroRNA-185; Proliferation; VEGFA; VHL