[双靶点反义RNA抑制乙型肝炎的实验研究]

[Experimental study of inhibition of hepatitis B by dual-target antisense RNA].

作者信息

Wu C, Zeng Z, Wang Q

机构信息

Department of Infectious Diseases, First Hospital, Peking University, Beijing 100034, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2001 May 25;81(10):605-8.

PMID:11798933

Abstract

OBJECTIVE

To investigate the effect of retroviral vector expressing the dual--target antisense RNA of hepatitis B virus (HBV) on the replication and expression of HBV.

METHODS

Recombinant retroviral vector plasmids expressing dual-target antisense RNA complementary to HBV X (1400 - 1430) and P (2375 - 2405) were constructed and transduced into 2.2.15 cells. The experimental cells were divided into PLXSN + X, PLXSN + P, PLXSN + X/P, and PLXSN + Xpos/P pos groups. The HBV expression was tested by ELISA method. HBV DNA and RNA were tested by FQ-PCR.

RESULTS

The inhibition rates of 2.2.15 cells transduced with recombinant vector plasmids (PLXSN + X group, PLXSN + P group, especially the PLXSN + X/P group) on expression of HBV DNA and RNA were higher than those of blank control group, PLXSN group, and PLXSN + Xpos/Ppos group. The expression of the three ORFs, S, C, and P were significantly inhibited.

CONCLUSION

The expression of HBV antisense RNA in 2.2.15 cells can inhibit the replication and expression of HBV. The inhibitory effect of dual antisense-RNA group is higher than that of single antisense-RNA group. Dual antisense RNA has the potentiality in anti-HBV gene therapy.

摘要

目的

研究表达乙型肝炎病毒（HBV）双靶点反义RNA的逆转录病毒载体对HBV复制和表达的影响。

方法

构建表达与HBV X（1400 - 1430）和P（2375 - 2405）互补的双靶点反义RNA的重组逆转录病毒载体质粒，并转导至2.2.15细胞中。将实验细胞分为PLXSN + X、PLXSN + P、PLXSN + X/P和PLXSN + Xpos/P pos组。采用ELISA法检测HBV表达。采用荧光定量聚合酶链反应（FQ-PCR）检测HBV DNA和RNA。