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Experimental study of TGF-β1-induced human hepatocytes line HL7702 epithelial-mesenchyme transition |
(School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang Jiangsu 212013, China) |
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Abstract Objective: To investigate whether hepatocytes could undergo epithelial-mesenchyme transition(EMT)on recombinant TGF-β1 stimulation in vitro . Methods: Human hepatocyte line HL7702 were cultured for 2 days with 20% NBS(1640)nutrient medium,and changed for serum-free 1640 medium and TGF-β1 factor induced HL7702 cells for 24,48,72 h. Cells morphology changes were watched before and after induction with inverted microscope. Real-time RT-PCR and western blot were used to analyze the expression of epithelial and mesenchyme cells-associated genes and proteins. Results: Hepatocytes before TGF-β1 induction were irregular sixangle cells. After stimulation cells showed spread and spindle-shaped morphology and cell-cell junction got bigger. Real-time RT-PCR and western blot analysis showed that epithelial marker-associated E-cadherin expression decreased obviously after TGF-β1 induction,while mesenchyme markers N-cadherin、Vimentin、Twist increased significantly. Conclusion: Human hepatocyte line HL7702 happened to EMT by recombinant TGF-β1 stimulation. EMT associated epithelial and mesenchyme genes and proteins expression changed significantly. It could be served as a foundation for further investigation on the mechanism of hepatocytes EMT.
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