人剪切修复基因XPD对p52和p21基因的影响  

Biological Behavior of HepG2 Hepatoma Cells Transfected by XPD in Vitro

作　　者：马果果[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》2011年第6期490-492,577,共4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:探讨人剪切修复基因XPD转染人HepG2肝癌细胞后p52和p21基因表达的变化以及对肝癌细胞生长的影响。方法:人肝癌细胞HepG2为靶细胞,XPD基因通过Lipofectamine2000脂质体转染HepG2。将细胞分为重组质粒HepG2-pEGFP-N2-XPD组(XPD组)、空载质粒HepG2-pEGFP-N2组(N2组)和HepG2空白对照组。分别用逆转录聚合酶链反应(RT-PCR)和蛋白印迹(Westernblot)法检测各组细胞中XPD、p52以及p21的mRNA和蛋白质的表达量,并用四甲基偶氮唑盐微量酶反应比色(MTT)法检测各组细胞的增殖能力。结果:XPD组中的p52的mRNA表达较其他2组显著下调,XPD、p21的mRNA表达较其他2组明显上调(均P<0.01)。Westernblot检测结果显示各组细胞中XPD、p52及p21的蛋白表达各组间差异与其mRNA各组间差异一致。MTT检测显示XPD转染入HepG2后细胞增殖能力减弱。结论:XPD基因可以抑制癌细胞的生长和基因p52的表达,促进p21的表达。Objective：To investigate the changes in expression of p52 gene and p21 gene, and the biological effects on hepatoma cell HePG2 after transfection of XPD. Methods： Targeting HepG2, the XPD gene was transfected into hepatoma cell HepG2 with Lipofectamine 2000. There were three groups in this study including HepG2-pEGFP-N2-XPD group（XPD group）, HepG2-pEGFP-N2 group （N2 group） and blank control group. The expressions of XPD, p52 and p21 were detected by RT-PCR and Western blot. The cell cycle was examined by MTT. Results： Compared with blank control group and N2 group , the expression of p52 mRNA decreased significantly in XPD group （P 〈 0.01）, but the expressions of XPD and p21 mRNA were enhanced obviously in XPD group （P 〈 0.01）. The trends of XPD, p52 and p21 protein were consistent with trend of mRNA detected by Western blot. MTT results showed that cell proliferation increased in XPD group. Conclusion： The wild-type XPD could inhibit the proliferation of HepG2 cells in vitro. The wild-type XPD could decrease the expression of p52 and enhance the expression of p21.

关 键 词：肝肿瘤 癌 NF-κB p52亚基 原癌基因蛋白质p21(ras) 逆转录聚合酶链反应 印迹法 蛋白质DNA修复 转染 XPD基因 

分 类 号：R730.21[医药卫生—肿瘤]