HBX蛋白对正常肝细胞增殖及端粒酶活性的影响  

The effect of HBX protein on the proliferation of normal hepatic cells and on telomerase activity

作　　者：贺丹丹[1] 邓志华[1] 杨倩[1] 

机构地区：[1]山西省太原市山西医科大学第二医院消化科,030001

出　　处：《癌变．畸变．突变》2012年第3期183-188,共6页Carcinogenesis,Teratogenesis & Mutagenesis

摘　　要：目的:观察乙型肝炎病毒X(hepatitis B virus X,HBX)蛋白对L02肝细胞的转化作用及其对端粒酶活性的影响。方法:构建PEGFP-N1-HBX质粒,脂质体介导转染L02细胞,倒置荧光显微镜观察EGFP的表达。稳定转染后Western blotting检测L02细胞中HBX蛋白的表达,电子显微镜观察L02细胞的形态,MTT法检测细胞的增殖,RT-PCR检测L02细胞端粒酶逆转录酶(hTERT)mRNA的表达,TRAP-PCR银染法检测L02细胞端粒酶活性的变化。结果:成功构建了PEGFP-N1-HBX质粒,稳定转染后L02细胞中有HBX蛋白的表达,且细胞形态明显幼稚化,有恶性变趋势,细胞增殖能力明显增加(P<0.05),细胞hTERT mRNA表达明显增加,端粒酶被激活。结论:HBX蛋白可诱导正常肝细胞的恶性转化,上调hTERT mRNA的表达并能激活端粒酶。OBJECTIVE：To observe the transformation in normal hepatic cell line L02 by the hepatitis B virus X（HBX） protein and the effect of HBX protein on the activity of telomerase.METHODS：PEGFP-N1-HBX plasmids were constructed and transfected into L02 normal liver cells,the expression of EGFP was assessed by fluorescence microscope.The expression of HBX protein in L02 cells was measured by Western blotting analysis.Cellular morphology was studied by electron microscope.The proliferation of L02 cells after stably transfected with plasmids was measured by MTT assay.The expressions of hTERT mRNA was detected by RT-PCR in L02 cells after stably transfected with plasmids.The activity of telomerase was evaluated by TRAP-PCR argentation in L02 cells after stably transfected with plasmids.RESULTS：PEGFP-N1-HBX plasmid was successfully constructed.The expression of HBX protein was observed in normal liver L02 cells.The cell morphology after transfection with PEGFP-N1-HBX plasmid appeared immature and with the trend of malignant transformation.Compared with the L02 cells after transfection with PEGFP-N1 plasmid,PEGFP-N1-HBX transfection promoted the proliferation of L02 cells（P〈0.05）.The hTERT mRNA expression increased in L02 cells after transfection with PEGFP-Nl-HBX plasmid and telomerase was activated.CONCLUSION： HBX protein could induce normal liver L02 cells torword malignant transformation,enhance the expression of hTERT mRNA and activate telomerase.

关 键 词：乙型肝炎病毒X蛋白 肝肿瘤 端粒酶 基因治疗 

分 类 号：R735.7[医药卫生—肿瘤]