The Effects Of XPD In Human HepG2Hepatoma Cells Regulating Rb And MAD2Genes

Objective：To investigate the role of xeroderma pigmentosum D(XPD)on the growth ofhuman HepG2hepatoma cells and the expressions of Rb and MAD2genes.Methods：1.Cultivated Human HepG2hepatoma cells in DMEM containing10%FBS.2.There combinant plasmid pEGFP-N2／XPD and empty plasmid pEGFP-N2were transfected into human HepG2hepatoma cells by lipofectamine2000TM；thereare4groups including blank control group,lipofectamine group,pEGFP-N2group（N2group）pEG FP-N2／XPD group（XPD group）3.Using TRIZOL reagent to extracted Total RNA from the cells,By usingRT-PCR analyze the mRNAexpression levels of XPD,Rb,MAD2genes.4.Total protein was extracted from the cells using total protein extraction kit,Theidentification of protein expression situation of XPD,Rb,MAD2genes by usingWestern blotting.5.Spread the cells in96-well plates,after transfection with plasmids,Cell growthwas detected by MTT.6.Using Flow cytometry(FCM)to analyze the cell apoptosis and cell cycle.Results：1.After transfection for24hour,the green fluorescent were covered over75%inpEGFP-N2and pEGFP-N2-XPD under fluorescebt microscope(×100)2.Compare with others,up-regulated the mRNA expression of XPD and Rb genein XPD group（P <0.05）but down-regulated the expression of MAD2gene（P <0.05）3、Compare with others, up-regulated the protein expression of XPD and Rbgene in XPD group（P <0.05）but down-regulated the expression of MAD2gene（P<0.05）4.MTT results showed that XPD restrain the proliferation of human HepG2hepatoma cells. 5.FCM showed that XPD exacerbated the apoptosis and prevented the hepatomacells from G1stage to S stage.Conclusions:XPD could restrain the growth of hepatoma cells,up-regulate the expression ofRb,and down-regulate the expression of MAD2.