title: MicroRNA-200a inhibits epithelial-mesenchymal transition in human hepatocellular carcinoma cell line.
abstract: OBJECTIVE
  Our study investigated the role of microRNA (miR)-200a and its molecular targets in hepatocellular carcinoma (HCC) cells.
  
  
  METHODS
  An inhibitor of miR-200a was transiently transfected into the hepatocellular carcinoma cell line, MHCC-97L. The effect of this transfection on mRNA levels of epithelial-mesenchymal transition (EMT)-related genes was measured by fluorescence-based quantitative real-time polymerase chain reaction (qRT-PCR). Further, protein levels of EMT-related genes, cell proliferation and apoptosis-related markers were assessed by Western blot analysis in these transfected cells. MTT and wound-healing assay were used to evaluate the proliferation and migration of MHCC-97L cells in presence and in absence of miR-200a inhibitor.
  
  
  RESULTS
  Compared with miR-NC control group, qRT-PCR results in anti-miR-200a group revealed a significant reduction in the mRNA levels of E-cadherin, with a concomitant increasing in vimentin mRNA level (all P < 0.05). Western blot results showed higher E-cadherin and Caspase-3 protein expressions in anti-miR-200a group compared to miR-NC group (P < 0.05). In addition, vimentin and Ki-67 protein expression was found sharply decreased in anti-miR-200a group compared to miR-NC group (P < 0.05). Consistent with this, wound-healing and MTT assay showed that migration and proliferation capacity of MHCC-97L cells in anti-miR-200a group is significantly increased compared with miR-NC group (both P < 0.05).
  
  
  CONCLUSIONS
  Our study reveals an important role of miR-200a in inhibiting EMT, proliferation and migration in HCC cells, suggesting the possibility of miR-200a-based therapeutics in HCC.
authors.affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine Guangzhou 510405, P. R. China.
authors.forename: Chong
authors.initials: C
authors.lastname: Zhong
authors.affiliation: Department of Radiotherapy, Cancer Center of Gangzhou Medical University Guangzhou 510095, P. R. China.
authors.forename: Ming-Yi
authors.initials: MY
authors.lastname: Li
authors.affiliation: Department of Gastrointestinal Surgery, The Sixth Affiliated Hospital of Sun Yat-Sen University Guangzhou 510655, P. R. China.
authors.forename: Zhi-Yuan
authors.initials: ZY
authors.lastname: Chen
authors.affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine Guangzhou 510405, P. R. China.
authors.forename: Hai-Kun
authors.initials: HK
authors.lastname: Cheng
authors.affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine Guangzhou 510405, P. R. China.
authors.forename: Ming-Li
authors.initials: ML
authors.lastname: Hu
authors.affiliation: Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine Guangzhou 510405, P. R. China.
authors.forename: Yue-Lu
authors.initials: YL
authors.lastname: Ruan
authors.affiliation: Department of Hepatobiliary Surgery, Sun Yat-Sen University Cancer Center Guangzhou 510060, P. R. China.
authors.forename: Rong-Ping
authors.initials: RP
authors.lastname: Guo
mesh_terms: D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D035683:MicroRNAs / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection
chemical_list: C533896:MIRN200 microRNA, human; D035683:MicroRNAs
keywords: MHCC-97L; MicroRNA-200a; epithelial-mesenchymal transition; hepatocellular carcinoma; migration; proliferation