人剪切修复基因XPD对人肝癌SMMC-7721细胞中Ets-1和Cdk6基因的调控作用  

Role of Excision Repair Gene XPD in Regulation of Ets-1 and Cdk6 of Hepatoma Cells

作　　者：王娟[1] 杜芳腾[1] 杜瑶[1] 张吉翔[1] 

机构地区：[1]南昌大学第二附属医院消化科、江西省分子医学重点实验室,330006

出　　处：《天津医药》2012年第9期865-869,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:30360037)

摘　　要：目的:观察野生型人剪切修复基因XPD转染入人肝癌细胞SMMC-7721后,细胞内Ets-1和Cdk6基因的表达变化及对SMMC-7721肝癌细胞增殖的影响。方法:将人工合成的pEGFP-N2-XPD重组质粒通过Lipofectamine 2000TM转染SMMC-7721细胞。设重组质粒转染细胞SMMC-7721-pEGFP-N2-XPD(XPD)组、空载质粒转染细胞SMMC-7721-pEGFP-N2(N2)组、脂质体组、无转染空白对照组。分别用逆转录-聚合酶链反应(RT-PCR)和蛋白印迹法(Western blot)检测细胞中XPD、Ets-1、Cdk6基因mRNA和蛋白质的表达量,并用流式细胞仪检测细胞周期变化,四甲基偶氮唑盐微量酶反应比色法(MTT)检测各组细胞的增殖活力。结果:XPD组中的XPD的mRNA和蛋白质表达较其他3组明显增高(P<0.001),而Ets-1、Cdk6 mRNA和蛋白质表达较其他3组明显减少(P<0.001)。转染pEGFP-N2-XPD重组质粒后细胞停滞在G1期,难于进入S期。转染了野生型XPD的SMMC-7721细胞增殖能力减弱。结论:XPD基因可能通过抑制Ets-1、Cdk6基因的表达影响肝癌细胞的生长。Objective： To observe the expressions of Ets-1 and Cdk6 gene and their effects on the growth and proliferation of SMMC-7721 cells transfected with wild-type XPD gene. Methods：The pEGFP-N2-XPD was transfected into SMMC-7721 cells by Lipofectamine 2000^（TM）. There were four groups in this study including SMMC-7721-pEGFP-N2-XPD （XPD） group,SMMC-7721-pEGFP-N2 （N2） group, lpofectamine （Lip） group and blank control group. The expression levels of XPD,Ets-1 and Cdk6 were detected by RT-PCR and Western blot. Flow cytometry （FGM） was used to analyze the cell cycle of SMMC-7721 cells. The cell proliferation was measured by MTI assay. Results： The expression of XPD mRNA and protein was significantly higher in XPD group than that of other three groups （P 〈 0.001）. The expressions of Ets-1 and Cdk6 mRNA and proteins were significantly decreased compared with those of other three groups （P 〈 0.001）. FCM result showed that XPD prevented the hepatoma ceils from G1 stage to S stage. The preliferation ability of SMMC-7721 cells was significantly reduced after transfetion with wild-type XPD gene （P 〈 0.01）. Conclusion： XPD gene may regulate the proliferation of hepatoma ceils by inhibiting Ets-1 and Cdk6 genes.

关 键 词：着色性干皮病蛋白质D组 肝肿瘤 原癌基因蛋白质c-ets-1 细胞周期蛋白依赖激酶6 转染细胞周期 

分 类 号：R282.710.5[医药卫生—中药学]