针对P基因的siRNA对乙肝病毒S表达稳定性的影响  

The inhibitory effect of HBs targeted by siRNA of HBV polymerase

作　　者：姚津剑[1] 林菊生[1] 韩思源[1] 徐冬[1] 于伟玲[2] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝病研究所,湖北武汉430030 [2]华中科技大学同济医学院附属同济医院肿瘤中心

出　　处：《胃肠病学和肝病学杂志》2007年第6期548-551,共4页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的以乙型肝炎病毒(HBV)P基因为靶点,构建表达小干扰RNA(siRNA)的质粒载体psRNA1、psRNA2、psRNA3、psRNA4及对照psiRNA0,体外观察针对P基因的siRNA对HBs表达的影响。方法设计并合成针对HBV-P基因的siRNA寡核苷酸,经退火形成双链后克隆入psiRNA-H1neo质粒中,通过鉴定将构建成功的5个psiRNAs真核表达质粒瞬时转染HepG2.2.15细胞,用半定量RT-PCR对筛选到的位点进行了试验观测抑制效果。结果表明针对P基因的siRNA能下调HBs的表达,其中psiRNA1、psiRNA2的抑制HBs基因表达的效果最强。结论针对P基因的siRNA可以有效的抑制HBs的表达,抑制的效果和siRNA的靶位点有关。Objective To construct psiRNA vectors expressing short interfering RNAs （siRNA） targeting hepatitis B virus （HBV） P gene sequence and to evaluate the inhibitory effect of HBs gene expression in HepG2.2. 15 cell. Methods Base on the sequence of HBV 2.2.15 cell, four sequences targeting HBV-P gene were designed and cloned into siRNA expression vector psiRNA-Hlneo. The constructed plasmids were transiently transfection into HepG2.2.15 cell. Result The plasmid-derived psiRNA1 and psiRNA2 could effectively inhibit transcription and translation of HBs gene（56.2% and55.3% ） , compared to other psiRNAs. Conclusion siRNA targeting against P gene could dwonregulate the expression of HBs, the inhibitory effect is majorly related to the targeted site of siRNA.

关 键 词：RNA干扰 小干扰RNA(siRNA) HBV聚合酶 

分 类 号：R512.62[医药卫生—内科学]