The Role Of XPD In Hepatoma Cell Growth And REDD1Gene Expression In Vitro

Objective:To investigate the effects of transfected XPD gene on the growth of hepatoma cell HepG2and the expression of REDD1gene in the cells in vitro.Methods:The recombinant plasmid pEGFP-N2/XPD and experimental plasmid pEGFP-N2through empty liposomes (Lipofectamine TM2000) mediated transient transfection into human hepatoma cell line HepG2. Experiment was divided into four groups, respectively, human hepatoma cell line HepG2(blank control group),liposome transfection human hepatoma cell line HepG2group (Lip group),empty plasmid transfection human hepatoma cell line HepG2-pEGFP-N2group (N2group),recombinant plasmid transfection human hepatoma cell line HepG2-pEGFP-N2-XPD group (XPD group). Fluorescence microscopy of green fluorescent protein gene expression, reverse transcription polymerase chain reaction (RT-PCR), Western blotting (Western blot) measured in each group XPD, REDD1gene mRNA,and protein expression levels. The cell was apoptosis measured by flow cytometry; The cell proliferation was detected by MTT.Results:Fluorescence microscopy to transfected with the recombinant plasmid pEGFP-N2/XPD and empty plasmid pEGFP-N2cells green fluorescence, indicating successful transfection. RT-PCR and Western blot showed that compared with other three groups, XPD group mRNA and protein expression levels of XPD, REDD1was significantly increased(P<0.01). The cell proliferation was obviously declined in XPD group (P<0.01);Flow cytometry showed that XPD group significantly increased apoptosis rate reached45%(P<0.01).Conclusion:XPD can increase the expression of tumor suppressor gene REDD1and could significantly inhibit the proliferation of HepG2cells induce apoptosis.