title: Interference of suppressor of cytokine signaling 3 promotes epithelial-mesenchymal transition in MHCC97H cells.
abstract: OBJECTIVE
  To investigate the role of suppressor of cytokine signaling 3 (SOCS3) silencing in epithelial-mesenchymal transition (EMT) involved in a human hepatocellular carcinoma MHCC97H cell line.
  
  
  METHODS
  MHCC97H cells were transiently transfected with SOCS3 small-interfering RNA (siRNA). Morphological changes of the transfected cells were observed under microscope. Expressions of E-cadherin, Vimentin and α-smooth muscle actin (α-SMA) were identified with immunofluorescence. Furthermore, protein expressions and mRNA levels of characteristic markers of EMT (E-cadherin, Vimentin, α-SMA and Snail) were detected by Western blotting, quantitative real-time polymerase chain reaction. Transforming growth factor-β1 (TGF-β1) levels in the supernatant were measured with enzyme-linked immunosorbent assay.
  
  
  RESULTS
  The transfected cells with SOCS3 siRNA showed a morphological alteration from a typical cobblestone morphology to mesenchymal spindle-shaped and fusiform features. SOCS3 siRNA lessened immunofluorescent expression of E-cadherin, but elicited immunofluorescent expressions of Vimentin and α-SMA in MHCC97H cells. More importantly, compared with the negative control, depletion of SOCS3 resulted in the decrease of the epithelial marker E-cadherin (P < 0.05), and the increase of the mesenchymal markers Vimentin and α-SMA and the transcription factor Snail in MHCC97H cells (P < 0.05). Moreover, compared with the negative control, SOCS3 siRNA evidently enhanced TGF-β1 secretion in MHCC97H cells (200.20 ± 29.02 pg/mL vs 490.20 ± 92.43 pg/mL, P < 0.05).
  
  
  CONCLUSIONS
  SOCS3 silencing is able to promote EMT in MHCC97H cells via changing the phenotypic characteristics and modulating the characteristic markers.
authors: Scientific Research Center, the Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Xi'an 710004, Shaanxi Province, China.
authors: Yuan-Yuan
authors: YY
authors: Ji
authors: Zhi-Dong
authors: ZD
authors: Wang
authors: Zong-Fang
authors: ZF
authors: Li
authors: Ke
authors: K
authors: Li
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D048430:Cell Shape; D058750:Epithelial-Mesenchymal Transition; D005838:Genotype; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D010641:Phenotype; D034622:RNA Interference; D015398:Signal Transduction; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; C106644:SOCS3 protein, human; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1
keywords: E-cadherin; Epithelial-mesenchymal transition; Hepatocellular carcinoma; Snail; Suppressor of cytokine signaling