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Effect of human xeroderma pigmentosum D on expression of Mdm2 and Mdm4 in HepG2 cells
Authors:DING Hao  ZHANG Ji-xiang
Affiliation:Department of Gastroenterology, the Second Affiliated Hospital of Nanchang University, Nanchang 330006, China.
Abstract:AIM:To investigate the effects of human xeroderma pigmentosum D (XPD) on the expression of murine double minute 2 (Mdm2) and murine double minute 4 (Mdm4) in human hepatoma cells. METHODS:Recombinant plasmid pEGFP-N2/XPD and vacant plasmid pEGFP-N2 were transfected into HepG2 cells using liposome, and the cells were divided into blank control group, pEGFP-N2 group and pEGFP-N2/XPD group. The cell growth was detected by MTT assay. The cell cycle and apoptotic rate were examined by flow cytometry. The mRNA and protein expression levels of XPD, Mdm2, Mdm4 and P53 were determined by RT-PCR and Western blotting. RESULTS:The results of MTT assay showed that the cell growth was inhibited by the transfection of pEGFP-N2/XPD. The results of flow cytometry showed that the transfection of pEGFP-N2/XPD increased the cell number in G 1 phase, decreased the cell number in S phase and increased the apoptotic rate of HepG2 cells. The results of RT-PCR and Western blotting showed that the transfection of pEGFP-N2/XPD increased the expression of XPD, decreased the expression of Mdm2 and Mdm4, and increased the expression of P53. CONCLUSION:XPD down-regulates Mdm2 and Mdm4 expression and up-regulates P53 expression in hepatoma cells. Moreover, the proliferation of hepatoma cells can be inhibited and the apoptosis can be induced by XPD.
Keywords:Xeroderma pigmentosum D  HepG2 cells  Murine double minute 2  Murine double minute 4  
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