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Silencing PPARγ gene inhibits proliferation and inducs apoptosis of hepatoma HCCLM3 cells

WU Xu-dong1,CHEN Wei-chang2,HUANG Xiao-ping2(1.Department of Gastroenterology,First People's Hospital of Yancheng City,Yancheng 224001,China;2.Department of Gastroenterology,First Affiliated Hospital of Soochow University,Suzhou 215006,China)  
Objective: To observe the effects of knocking down the expression of peroxisome proliferators-activated receptor gamma(PPARγ) with RNA interference techniques on the proliferation and apoptosis of hepatocellular carcinoma HCCLM3 cells.Methods:A short-hairpin RNA(shRNA) eukaryotic expression vector against PPARγ was constructed and transfected into HCCLM3 cells.The changes of PPARγ expression were detected by semi-quantitative RT-PCR and Western blotting analysis.The proliferation of HCCLM3 cells was tested by MTT assay.Apoptosis ratio of HCCLM3 cells was detected by TUNEL method and flow cytometry(FCM).Expressions of PCNA and wide-type p53 protein were analyzed by immuocytochemistry(ICC) methods.Results:The sequence-specific shRNA(pshPPARγ) efficiently blocked the expression of PPARγ mRNA by 80.5%.At 48 h after transfection of pshPPARγ,proliferation of HCCLM3 cells was significantly suppressed by 71.5%.The positive rate of PCNA expression was(23.8±7.2)% at 40 h transfection.The apoptotic rates were(24.2±4.7) % as detected by TUNEL assay and(23.2±4.2)% of cells as measured by FCM test,respectively.The detection results of the two methods were consistent.In pshPPARγ transfection group,cell cycle of HCCLM3 cells was arrested in G0/G1 phase and the proportion of cells in G2/M phase decreased.Moreover,expression of wide-type p53 protein increased significantly.Conclusion:Knockdown PPARγ expression with RNA interference technology can significantly suppress proliferation and induce apoptosis of HCCLM3 cells.It is related with up-regulation of wide-type p53 protein expression.
【CateGory Index】: R735.7
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