丙型肝炎病毒1b基因型C区氨基酸70替代对细胞凋亡的影响  

Effects of the Amino Acid 70 Substitution in the Core Region of Hepatitis C Virus Genotype 1b on Cell Apoptosis

作　　者：胡中杰[1] 刘颖[1] 仇丽霞[1] 范作鹏[1] 聂巍[1] 梁珊[1] 于红卫[1] 金荣华[1] 

机构地区：[1]首都医科大学附属北京佑安医院丙肝与中毒性肝病科,100069

出　　处：《医学研究杂志》2014年第6期49-52,共4页Journal of Medical Research

基　　金：北京市优秀人才培养基金资助项目(2010D003034000009);中国初级保健基金会佑安肝病艾滋病基金资助项目(BJYAH-2011-073);首都医科大学基础-临床科研合作基金资助项目(1000172053-11JL61)

摘　　要：目的探讨丙型肝炎病毒(hepatitis C virus,HCV)1b基因型(genotype 1b,GT-1b)C区氨基酸(amino acid,AA)70替代对细胞凋亡的影响。方法利用前期构建的HCV GT-1b C区aa70野生型和变异型核心蛋白表达质粒瞬时转染体外培养肝瘤细胞系,采用PCR芯片检测细胞凋亡相关基因的表达,并以Annexin-V FITC/PI双染色法检测细胞凋亡率。结果与野生型核心蛋白相比,转染表达变异型核心蛋白可使Huh7细胞84个凋亡相关基因中40个表达相对增强,44个表达相对减弱,但相差倍数均在3倍以内。HepG2细胞转染野生型和变异型核心蛋白表达质粒后,细胞凋亡率分别为9.4%±1.9%、9.1%±2.2%,低于空白载体对照组的14.3%±1.4%;以抗Fas抗体处理后,细胞凋亡率增高(52.4%±2.9%、56.1%±2.5%),但仍明显低于空白载体对照组的72.6%±2.1%。结论 HCV核心蛋白表达可抑制细胞凋亡,但核心蛋白单纯R70Q/H变异并不足引起细胞凋亡相关基因表达及凋亡率的显著不同。Objective To investigate the effects of the amino acid （aa） 70 substitution in the core region （CR） of hepatitis C virus （HCV） genotype l b （GT- 1 b） on cell apoptosis. Methods The expression plasmids of wild type and mutant aa70 core proteins weretransfected into human hepatoma cell line. Apoptosis PCR array was performed to test the expression of 84 key genes involved in programmed cell death. And then, apoptosis rate was measured by flow cytometry （FCM）. Results Compared with HCV aa70 wild type core protein, expression of mutant core protein in Huh7 cells could make 40 genes＇ expression increased and 44 genes＇ expression decreased among the 84 key genes related to apoptosis. However, the difference of multiples was within 3 times. Furthermore, when transfected with HCV aa70 wild type or mutant core protein expression plasmids, the apoptosis rates of HepG2 cells were 9.4% ± 1.9% and 9.1% ± 2.2% respectively, which were lower than those in control group transfected with empty vector （ 14.3% ± 1.4% ）. Moreover, after treated with anti - Fas antibody, the apoptosis rates of HepG2 cells could rise to 52.4% ± 2.9% and 56.1% ±2.5% respectively, which was still lower than that in control group （72.6% ± 2.1% ）. Conclusion The expression of HCV core protein can inhibit fas - mediated apoptosis. But RTOQ/H substitution alone is not sufficient for the significantly difference of the expression of apoptosis related genes and the cells＇ apoptosis rates.

关 键 词：丙型肝炎病毒 核心蛋白 凋亡 

分 类 号：R5[医药卫生—内科学]