人剪切修复基因XPD对肝癌细胞P53和Ets-1基因的作用  

The role of excision repair gene XPD in regulation of P53 and Ets- 1 in hepatoma cells

作　　者：田明[1] 游春娇 张吉翔[1] 

机构地区：[1]南昌大学第二附属医院消化内科,南昌330006 [2]南昌大学第二附属医院心内科,南昌330006

出　　处：《广东医学》2014年第8期1149-1153,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:30360037)

摘　　要：目的探讨人剪切修复基因XPD对肝癌细胞P53和Ets-1基因的影响。方法使用脂质体瞬时转染技术,将pEGFP-N2-XPD重组质粒转染至肝癌细胞HepG2,24 h后加入20μmol/L的P53抑制剂Pifithrin-α,孵育24 h,并以未经转染的HepG2细胞作为空白对照。RT-PCR、Western blot检测细胞中XPD、P53、phosphoP53(ser-15)、Ets-1的mRNA和蛋白的表达变化,MTT法观察细胞增殖的活力,流式细胞仪检测细胞周期变化。结果与空白对照比较,转染pEGFP-N2-XPD重组质粒后,HepG2细胞XPD、P53 mRNA和蛋白表达上调(P<0.01),而Ets-1 mRNA和蛋白表达下调(P<0.01),加入Pifithrin-α后,XPD、P53 mRNA和蛋白表达下调,Ets-1mRNA和蛋白表达上调(P<0.01)。转染pEGFP-N2-XPD重组质粒后,HepG2细胞进入S期发生阻滞,停滞在G1期(P<0.01),加入Pifithrin-α后,HepG2细胞G1期细胞减少,S期细胞增多(P<0.01)。转染pEGFP-N2-XPD重组质粒后,HepG2细胞增殖活力下降(P<0.01),加入Pifithrin-α后,HepG2细胞增殖活力上升(P<0.01)。结论野生型XPD基因在转染至肝癌HepG2细胞后,可以上调P53的表达,通过P53途径抑制Ets-1的表达,促使肝癌细胞凋亡。Objective To investigate the effects of xeroderma pigmentosum D（ XPD） on the growth of hepatoma cells and the expressions of P53 and Ets- 1. Methods Human hepatoma cells（ HepG2） were transfected with the plasmids of pEGFP- N2- XPD using Lipofectamine 2000,and subsequently incubated with 20 μmol / L Pifithrin- α（ P53 inhibitor） for 24 h. Blank control group（ no transfecting and adding medicine） was set. The expressions of XPD,P53, phospho- P53（ ser- 15） and Ets- 1 were assessed by RT- PCR and Western blot. Flow cytometry was used to examine the cell cycle and apoptosis. MTT assay was used to detect the cell proliferation. Results After transfected with the pEGFP- N2- XPD plasmid,the mRNA and protein expression of XPD and P53 were significantly up- regulated,while the expression of Ets- 1 was significantly down- regulated（ P〈0. 01）. Moreover,the cell proliferation was inhibited and the apoptosis was accelerated with pEGFP- N2- XPD plasmid transfected,which was reversed by Pifithrin- α. Conclusion Wild- XPD gene can restrain the proliferation,down- regulate Ets- 1 via P53 pathway,and impel the apoptosis of hepatoma cells.

关 键 词：肝肿瘤 人着色性干皮病D组基因 转染 P53基因 原癌基因蛋白质Ets-1 

分 类 号：R735.7[医药卫生—肿瘤]