靶向HBVs和e抗原基因的shRNA表达载体共转染对HBV抗原表达的抑制作用

Inhibition of HBsAg and HBeAg expression by shRNA expressing vectors targeting HBVs and egene

目的:探索靶向乙型肝炎病毒HBsAg基因的shR-NA表达载体Pgs1、Pgs2、Pgs3及靶向HBeAg基因shRNA表达载体psiHBV4、psiHBV5、psiHBV6共转染,对体外培养HepG2.2.15细胞中的HBV抗原表达的抑制作用。方法:以质粒PTZ为阴性对照,将自行构建的靶向HBVs和e抗原基因的shRNA表达载体按不同组合共转染HepG-2.2.15细胞;继续培养24 h后用MEIA分别检测细胞裂解液和培养上清中HBsAg和HBeAg的表达水平。结果:psiHBV4+PgS2、psiH-BV4+PgS3在联合转染时,在细胞上清和裂解液中对HBsAg和HBeAg表达都有显著抑制作用(P<0.05),psiHBV5+PgS1、psiHBV6+PgS3对裂解液HBeAg表达抑制作用不显著(P>0.05)。结论:靶向HBs和HBe基因的两种载体psiH-BV4+PgS2、psiHBV4+PgS3共转染比单个载体转染更能显著减少HBV抗原的表达。

AIM： To constructed the shRNA expressing vectors targeting HBsAg gene and HBeAg gene of HBV,Pgs1,Pgs2,Pgs3 and psiHBV4,psiHBV5,psiHBV6 in order to probe their inhibitation on the HBV antigen expression in outlive HepG2.2.15 cells.METHODS： PTZ was used as negative control,both the shRNA expressing vectors targeting HBsAg and HBeAg gene of HBV was together transfected into HepG-2.2.15 cells with different combinations,and detected the expressional level of the HBsAg and HBeAg in the cell disruption liquid and the cultivated supernatant with MEIA after 24 h.RESULTS： The group transfecting psiHBV4 and PgS2,psiHBV4 and PgS3 could significantly inhibit HBsAg and HBeAg expression compared with control group（P0.05）in the cell disruption liquid and the cultivated supernatant.But the group transfecting psiHBV5 and PgS1,psiHBV6 and PgS3 cannot significantly inhibit HBeAg expression（P0.05）in the cell disruption liquid.CONCLUSION： The shRNA expressing vectors targeting HBsAg and HBeAg gene of HBV psiHBV4 and PgS2,psiHBV4 and PgS3 could significantly inhibit the antigen expression of HBV than only one.