Viral Hepatitis
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jan 28, 2005; 11(4): 498-502
Published online Jan 28, 2005. doi: 10.3748/wjg.v11.i4.498
Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro
Zheng-Gang Yang, Zhi Chen, Qin Ni, Ning Xu, Jun-Bin Shao, Hang-Ping Yao
Zheng-Gang Yang, Zhi Chen, Qin Ni, Ning Xu, Jun-Bin Shao, Hang-Ping Yao, Institute of Infectious Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Zhi Chen, Institute of Infectious Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China. chenzhi@zju.edu.cn
Telephone: +86-571-87236579 Fax: +86-571-87068731
Received: December 12, 2003
Revised: December 18, 2003
Accepted: February 1, 2004
Published online: January 28, 2005
Abstract

AIM: To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA) expression vector.

METHODS: Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vector and shRNA expression vectors were constructed and cotransfected transiently into HepG2 cells. mRNAs extracted from HepG2 cells were detected by real-time PCR. Fluorescence of HBs-GFP protein was detected by fluorescence-activated cell sorting (FACS). The effective shRNA expression vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in HepG2.2.15 cells were analyzed by radioimmunoassay (RIA) method.

RESULTS: FACS revealed that shRNA targeting at HBsAg reduced the GFP signal by 56% compared to the control. Real-time PCR showed that HBs-GFP mRNA extracted from HepG2 cells cotransfected with pAVU6+27 and HBs-GFP expression plasmids decreased by 90% compared to the empty vector control. The expressions of HBsAg and HBeAg were also inhibited by 43% and 64%, respectively.

CONCLUSION: RNAi using shRNA expression vector can inhibit the expression of HBsAg, providing a fresh approach to screening the efficient small interfering RNAs (siRNAs).

Keywords: Hepatitis B Surface Antigens; Small hairpin RNA; RNA interference; Gene expression