肝肿瘤论文人剪切修复基因XPD对p52及p21基因的影响.docVIP

肝肿瘤论文：人剪切修复基因XPD对p52及p21基因的影响 【中文摘要】探讨人剪切修复基因XPD转染入HepG2盯癌细胞后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PCR(RT-PCR)和蛋白印迹(Western blot)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和p21的mRNA表达较其他2组显著升高(均P0.O1)。Western blot法检测显示在XPD组中p52的蛋白表达较N2组和空白对照组显著下降,而基因XPD及p21的蛋白表达较N2组和空白对照组显著升高(均P0.01)。细胞增殖活力检测(MTT)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。 【英文摘要】:To investigate the biological effects of the trans fected gene XPD on hepatoma cell HepG2.Methods: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 level of p52 mRNA declined significantly in XPD group (P 0.01);but the expressions levels of XPD and p21 mRNA were elevated obviously in XPD group(P0.01). The trends of XPD, p52 and p21 protein detected by Western blot were consistent with trend of mRNA. MTT results showed that cell proliferation increased in XPD group.Conclusions:The wild-type XPD could inhibit the proliferation of HepG2 cells in vitro. The wild-type XPD could decrease the expression of p52 and increase the expression of p21. 【关键词】肝肿瘤 癌 NF-κB，p52亚基 原癌基因蛋白质p22(ras) DNA修复 【英文关键词】liver neoplasms neoplasms nf-kappa b p52 subunit proto-oncogene proteins p21(ras) DNA repair 【目录】人剪切修复基因XPD对p52及p21基因的影响 摘要 3-4 ABSTRACT 4 第1章 前言 6-8 第2章 实验材料及主要方法 8-16 2.1 材料 8-9 2.1.1 细胞及质粒 8 2.1.2 主要实验试剂 8-9 2.1.3 主要设备仪器 9 2.2 实验方法 9-16 2.2.1 HepG2细胞培养 9-10 2.2.2 转染XPD基因入HepG2 10-11 2.2.3 RT-PCR分析 11-12 2.2.4 蛋白印迹法 12-14 2.2.5 测定细胞增殖活力 14-15 2.2.6 统计学分析 15-16 第3章 实验结果 16-20 3