title: RhoC is a major target of microRNA-93-5P in epithelial ovarian carcinoma tumorigenesis and progression.
abstract: BACKGROUND
  An increasing amount of evidence has revealed that microRNAs regulate various biological processes, including cell differentiation, cell proliferation, apoptosis, drug resistance, and fat metabolism. Studies have shown that miR-93's targetome in cancer has not been fully defined. Moreover, the role of miR-93 in epithelial ovarian carcinoma (EOC) remains largely unknown.
  
  
  METHODS
  MIR-93 mRNA expression in normal ovarian tissue, benign tumors, borderline tumors, primary ovarian carcinomas, and metastatic omentum was quantified. The ovarian carcinoma cell lines OVCAR3, SKOV3/DDP, and HO8910-PM were transfected with miR-93-5P, after which cell phenotype and expression of relevant molecules were assayed. Dual-luciferase reporter assay and a xenograft mouse model were used to examine miR-93 and its target gene RHOC (Ras homolog gene family member C).
  
  
  RESULTS
  MIR-93 mRNA expression was significantly lower in ovarian carcinomas and borderline tumors than in normal ovarian tissues (p < 0.05), and was lower in metastatic omentum than in relative primary ovarian carcinomas (p < 0.05). MIR-93 mRNA expression was also negatively associated with differentiation (well vs. poor and moderate) and International Federation of Gynecology and Obstetrics staging (FIGO stage I/II vs. stage III/IV) in ovarian carcinoma (p < 0.05), besides, miR-93 was higher expressed in mucinous adenocarcinoma than the other types (p < 0.05). MiR-93-5P overexpression reduced proliferation (p < 0.05); promoted G1 or S arrest and apoptosis (p < 0.05); suppressed migration and invasion (p < 0.05); and reduced RhoC, P70S6 kinase, Bcl-xL, matrix metalloproteinase 9 (MMP9) mRNA or protein expression; conversely, it induced P53 and cleaved PARP expression (p < 0.05). Dual-luciferase reporter assay indicated that miR-93 directly targeted RhoC by binding its 3' untranslated region. MiR-93-5P transfection also suppressed tumor development and RhoC expression (determined by immunohistochemistry) in vivo in the xenograft mouse model (p < 0.05).
  
  
  CONCLUSIONS
  This is the first demonstration that miR-93-5P may inhibit EOC tumorigenesis and progression by targeting RhoC. These findings indicate that miR-93-5P is a potential suppressor of ovarian cellular proliferation. The involvement of miR-93-5P-mediated RhoC downregulation in inhibiting EOC aggressiveness may provide extended insight into the molecular mechanisms underlying cancer aggressiveness.
authors: Department of Gynecology, The First Affiliated Hospital of China Medical University, Shenyang, 110001, P. R. China. ccllnm@163.com.
authors: Xi
authors: X
authors: Chen
authors: Department of Gynecology, The First Affiliated Hospital of China Medical University, Shenyang, 110001, P. R. China. chenshuo077003@163.com.
authors: Shuo
authors: S
authors: Chen
authors: Department of Gynecology, The First Affiliated Hospital of China Medical University, Shenyang, 110001, P. R. China. xiuyinling@126.com.
authors: Yin-Ling
authors: YL
authors: Xiu
authors: Department of Gynecology, The First Affiliated Hospital of China Medical University, Shenyang, 110001, P. R. China. sunkx90@163.com.
authors: Kai-Xuan
authors: KX
authors: Sun
authors: Department of Biochemistry and Molecular Biology, College of Basic Medicine, China Medical University, Shenyang, 110001, P. R. China. zongzhi_999@163.com.
authors: Zhi-Hong
authors: ZH
authors: Zong
authors: Department of Gynecology, The First Affiliated Hospital of China Medical University, Shenyang, 110001, P. R. China. yida.zhaoyang@163.com.
authors: Yang
authors: Y
authors: Zhao
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D063646:Carcinogenesis / Q000235:genetics* / Q000473:pathology; D002277:Carcinoma / Q000235:genetics* / Q000473:pathology; D059447:Cell Cycle Checkpoints / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D018450:Disease Progression; D005260:Female; D059585:G1 Phase Cell Cycle Checkpoints / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D055815:Young Adult; D020741:rho GTP-Binding Proteins / Q000235:genetics*; D000075024:rhoC GTP-Binding Protein
chemical_list: D020413:3' Untranslated Regions; C543350:MIRN93 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D020780:Matrix Metalloproteinase 9; C066690:RHOC protein, human; D020741:rho GTP-Binding Proteins; D000075024:rhoC GTP-Binding Protein
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