靶向HBV C基因的siRNA在HepG2.2.15细胞中对HBV的抑制作用  

Inhibitory effect of targeting C gene siRNA on hepatitis B virus in HepG 2.2.15 cells

作　　者：肖安[1] 刘斌[1] 焦晔[2] 赵欣[2] 刘明秋[2] 严维耀[2] 郑兆鑫[2] 

机构地区：[1]成都军区昆明总医院传染科,昆明650032 [2]复旦大学生命科学院基因工程国家重点实验室

出　　处：《西南国防医药》2012年第12期1277-1280,共4页Medical Journal of National Defending Forces in Southwest China

基　　金：国家自然科学基金(30972629)

摘　　要：目的研究靶向HBV C基因的短发夹状RNA(shRNA)表达质粒对HepG2.2.15细胞中HBV的抑制作用。方法设计构建两个靶向HBV C基因的shRNA表达质粒S1和S2,以及用于对照的非同源shRNA表达质粒S3。转染HepG2.2.15细胞后,通过RT-PCR和ELISA方法分别检测细胞中HBV C基因表达和细胞上清中HBV DNA、HBsAg和HBeAg表达。结果在转染后24 h时,HBV C基因mRNA表达量下降58%～83%,HBV DNA下降了2.44～3.36个log值,HBsAg和HBeAg的表达水平分别降低了51%～79%和50%～77%。S1、S2联合运用可以提高对HBV的抑制作用,而S3无抑制效果。结论靶向HBV C基因的shRNA表达质粒S1、S2及其联合运用,均能有效地抑制HepG2.2.15细胞中HBV的复制和表达。Objective To study the inhibitory effect of shRNA expression plasmid of HBV C gene on HBV in HepG 2.2.15 cells.Methods Two shRNA expression plasmids（S1 and S2） of targeting HBV C gene and one nonhomologous shRNA expression plasmid（S3） used for the control were designed and constructed.After the transfection of HepG 2.2.15 cell,the HBV C gene expression and expressions of HBV,DNA,HBsAg,and HBeAg in the supernatant were respectively detected by Real Time PCR and ELISA methods.Results After the transfection for 24 h,the mRNA expression of HBV C gene significantly decreased by 58%-83%;the expression of HBV DNA decreased by 2.44-3.36 log values;the expression levels of HBsAg and HBeAg decreased by 51%-79% and 50%-77%,respectively.Combined application of S1 and S2 could enhance the inhibitory effect on HBV,while S3 had no such effect.Conclusion The shRNA expression plasmids S1 and S2 and the combined application of both two can effectively inhibit the replication and expression of HBV in HepG 2.2.15 cells.

关 键 词：乙型肝炎病毒 RNA干扰 小干扰RNA 短发夹状RNA 

分 类 号：R512.6[医药卫生—内科学]