IPS-1 SNP在调控HBV感染后干扰素应答机制的研究  

Study on the function of IPS-1 polymorphisms in the mechanism of regulating interferon response in HBV infection

作　　者：刘柯慧[1] 汤伟亮[1] 项晓刚[1] 赖荣陶[1] 李凤棣 赵钢德[1] 吴海清[1] 谢青[1] 王晖[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院感染科,200025

出　　处：《肝脏》2014年第9期664-668,共5页Chinese Hepatology

基　　金：国家自然科学基金(编号:81070334);十二五国家科技重大专项资助项目(编号:2012ZX10005004-002);上海市公共卫生人才培养计划(GWDTR201202);上海市卫生和计划生育委员会重点课题(20134004);上海市科委科技支撑项目(13401902900)

摘　　要：目的探讨干扰素β启动子刺激因子1(IPS-1)不同单核苷酸多态性(SNP)基因型对HBV感染后干扰素应答作用机制。方法利用IPS-1野生型和rs17857295、rs7262903、rs7269320三种SNP基因型慢病毒表达载体,转染目的细胞HepG2、HepG2.2.15细胞,qPCR检测HBV感染和IPS-1不同SNP基因型对目的基因表达情况的影响,检测细胞转染后IFN-βmRNA表达水平,探讨IPS-1不同SNP基因型对HBV感染后细胞干扰素应答的机制。结果 IPS-1rs17857295和rs7262903两种SNP基因型转染HepG2.2.15细胞后,IPS-1基因表达水平显著低于野生型转染组(451.77比712.53,498.71比712.53,P<0.01);但IFN-βmRNA表达水平显著高于野生型转染组(分别为5.33比0.98和3.59比0.98,P<0.01);rs7269320 SNP基因型过表达后,IPS-1基因表达水平显著低于野生型转染组(290.63比712.53,P<0.01),但IFN-βmRNA表达水平无明显变化(0.74比0.98)。另外,rs17859295基因型分别转染HepG2和HepG2.2.15细胞后,后者IFN-βmRNA表达水平显著高于前者(5.33比2.25,P<0.01)。结论 IPS-1 rs17857295和rs7262903两种SNP基因型HBV感染者,尤其rs17857295 SNP基因型,清除病毒的能力可能强于野生型的感染者;而IPS-1野生型和rs7269320 SNP基因型HBV感染者可能更易形成病毒感染的慢性化。Objective To investigate the function of interferon-βpromoter stimulator-1 （IPS-1 ）polymorphisms in regulating interferon response in HBV infection and to provide experimental evidence for clarifying the antiviral immune mechanisms of chronic hepatitis B. Methods HepG2 and HepG2.2.15 were cultured in 6 well plates in DMEM media for 24 h. Lipofectamine 2000 was used to transfect different expression vectors of IPS-1 into HepG2 or HepG2.2.15 cells,such as wild type,rs17857295,rs7262903 and rs7269320,according to the manufacture instructions. After 48 h,IPS-1 levels in these cells were detected by real-time quantitative PCR to evaluate the influence of HBV infection or IPS-1 SNP polymorphisms on the expression level of IPS-1 gene. Furthermore,IFN-βlevels were detected by real-time quantitative PCR to explore the role of IPS-1 SNP polymorphisms in the mechanism of regulating interferon response in HBV infection. Results In HepG2.2.15 cells,which were transfected by expression vectors of IPS-1 rs17857295 or rs7262903 SNP genotypes,IPS-1 gene expression level was significantly lower than that in wild-type tranfection group （451.77 vs 712.53, P〈0.0001）（498.71 vs 712.53,P= 0.0002）,but expression level of IFN-βgene was higher than in wild-type tranfection group （5.33 vs 0.98）（3.59 vs 0.98）（P〈0.0001）. Otherwise,in rs7269320 SNP genotype transfected HepG2.2.15 cells group,IPS-1 gene expression level was significantly lower than that in wild-type tranfection group （290.63 vs 712.53,P〈0.0001）,but expression level of IFN-βgene didn’t have significant changes （0.74 vs 0.98）. Furthermore,compared with HepG2 cells,after expression vector of IPS-1 rs17857295 SNP genotype transfected HepG2 or HepG2.2.15 cells,IFN-βgene expression level in HepG2.2.15 cells was significantly higher after transfection of vector of IPS-1 rs17857295 SNP genotype （5.33 vs 2.25,P= 0.0005）. Conclusions Our study indicated that people with IPS-1 rs17857295 and rs7262903 SNP genotype,especially rs17857295 SNP

关 键 词：干扰素β启动子刺激因子-1 单核苷酸多态性 乙型肝炎病毒 基因定点突变 慢病毒 

分 类 号：R512.62[医药卫生—内科学]