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Effects of shRNA targeting SIAH2 gene on proliferation and apoptosis of human hepatoma cell line HepG2
Authors:XIAO Hai  HE Xing-bo  HUANG Cai-bin  LIU Yao
Affiliation:1.Department of Pathology, 2Department of Medical Services of the First Affiliated Hospital, 3Department of Gastroenterology of the First Affiliated Hospital, Gannan Medical College, Ganzhou 341000, China.
Abstract:AIM:To investigate the effects of shRNA-mediated seven in absentia homolog 2 (SIAH2; one of ubiquitin ligases) gene silencing on cell cycle and apoptosis of human hepatoma cell line HepG2. METHODS:The specific recombinant vector pGenesil-SIAH2 was transiently transfected into HepG2 cells with Lipofectamine 2000. RT-PCR and Western blotting were performed to detect the mRNA and protein expression levels of SIAH2. MTS assay was employed to evaluate the cell proliferation. Flow cytometry was used to determine the cell cycle and apoptosis of the transfected cells. RESULTS:Compared with control groups, the mRNA and protein levels of SIAH2 were reduced by pGenesil-SIAH2 transfection in HepG2-SIAH2 group. The proliferation of HepG2-SIAH2 cells was significantly inhibited. The percentage of G1-phase cells and the early apoptotic rate were significantly higher in HepG2-SIAH2 cells. CONCLUSION: Tansfection of pGenesil-SIAH2 effectively inhibits the proliferation of HepG2 cells, arrests the cells in G1 phase and induces apoptosis, indicating an experimental basis of SIAH2-targeting gene therapy for hepatocellular carcinoma.
Keywords:RNA interference  Ubiquitin ligases  HepG2 cells  Cell proliferation  Apoptosis  
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