不同基因型丙型肝炎病毒非结构蛋白NS3对端粒酶活性的研究  

Study of different genotypes hepatitis C virus non-structural 3 protein on telomerase activity

作　　者：李小宁[1] 杨正海[1] 李静[2] 

机构地区：[1]皖南医学院弋矶山医院检验科 [2]铜陵职业技术学院医学检验技术系

出　　处：《中国临床药理学与治疗学》2015年第2期128-131,共4页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：安徽高校省级自然科学研究项目(KJ2013B14)

摘　　要：目的:建立丙型肝炎病毒(HCV)NS3蛋白细胞表达模型,研究不同基因型丙型肝炎病毒端粒酶活性之间是否存在差异。方法:筛选出不同HCV基因型。分别运用PCR法扩增HCV NS3蛋白基因,转载真核表达载体pBK-CMV,构建重组质粒pBK-HCV NS3,分别导入肝癌细胞株HepG2,采用端粒重复序列扩增(TRAP)方法检测不同基因型HCV NS3蛋白基因转载质粒导入HepG2细胞的端粒酶活性。结果:本地区HCV基因型流行的主要是1b型,其次是2a型。转染不同基因型HCV NS3蛋白基因的HepG2细胞端粒酶活性较转染空载质粒的细胞明显升高;不同基因型HCV端粒酶活性之间存在差异,HCV 1b型的端粒酶活性明显高于HCV 2a型。结论:HCV NS3蛋白可能以某种机制激活了端粒酶活性。不同基因型HCV NS3蛋白在细胞恶性转化中的影响有差异。AIM：To establish an expression model of HCV NS3 protein and explore the differences of telomerase activities among different hepatitis C virus genotypes. METHODS：Screening of different HCV genotypes through PCR array.HCV NS3 gene fragments（HCV 1b NS3 and HCV 2a NS3） were amplified and cloned into eukaryotic protein expression vector pBK-CMV.Followed by the introduction of recombinant plasmids（pBK-HCV1 b NS3 and pBK-HCV2aNS3）and control vector into hepatoma cell lines HepG2,respectively.Telomerase activities of HepG2 cells were detected by telomeric repeat amplification protocol（TRAP）assay.RESULTS：The popular genotype of HCV in our region was HCV 1b,second was HCV 2a.Telomerase activities in HepG2 cells transfected with pBK-HCV NS3 significantly increased,compared with control. Furthermore,telomerase activities of HepG2 cells transfected with recombinant pBK-HCV1 b NS3 were higher as compare with the cells transfected with pBKHCV2a NS3 distinctly.CONCLUSION： HCV NS3 protein may activate telomerase through the same pathway.Different genotypes of HCV NS3 protein have different effects on the malignant transformation of HepG2 cells.

关 键 词：HCV NS3蛋白 基因型 端粒酶 

分 类 号：R446.5[医药卫生—诊断学]