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Journal of Virology, November 2005, p. 14392-14403, Vol. 79, No. 22
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.22.14392-14403.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Genomic Analysis of Anti-Hepatitis B Virus (HBV) Activity by Small Interfering RNA and Lamivudine in Stable HBV-Producing Cells

Yong Guo,1,2,4 Hongyan Guo,1,2,4 Liang Zhang,4 Hongying Xie,4 Xin Zhao,4 Fangxun Wang,4 Ze Li,4 Yahui Wang,4 Shiliang Ma,5 Jiaping Tao,5 Weixing Wang,6 Yuxiang Zhou,1,2,3,4,7 Weiping Yang,4,7 and Jing Cheng1,2,3,4,7*

Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China,1 Medical Systems Biology Research Center, Tsinghua University School of Medicine, Beijing 100084, China,2 The State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing 100084, China,3 National Engineering Research Center for Beijing Biochip Technology, 18 Life Science Parkway, Changping District, Beijing 102206, China,4 Analysis Center of Medicine, Pharmacy and Public Health, Peking University, Beijing 100083, China,5 The State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Science, Peking University, Beijing 100083, China,6 CapitalBio Corporation, 18 Life Science Parkway, Changping District, Beijing 102206, China7

Received 25 May 2005/ Accepted 24 August 2005

Hepatitis B virus (HBV) causes acute and chronic hepatitis and hepatocellular carcinoma. Small interfering RNA (siRNA) and lamivudine have been shown to have anti-HBV effects through different mechanisms. However, assessment of the genome-wide effects of siRNA and lamivudine on HBV-producing cell lines has not been reported, which may provide a clue to interrogate the HBV-cell interaction and to evaluate the siRNA's side effect as a potential drug. In the present study, we designed seven siRNAs based on the conserved HBV sequences and tested their effects on the expression of HBV genes following sorting of siRNA-positive cells. Among these seven siRNAs, siRNA-1 and siRNA-7 were found to effectively suppress HBV gene expression. We further addressed the global gene expression changes in stable HBV-producing cells induced by siRNA-1 and siRNA-7 by use of human genome-wide oligonucleotide microarrays. Data from the gene expression profiling indicated that siRNA-1 and siRNA-7 altered the expression of 54 and 499 genes, respectively, in HepG2.2.15 cells, which revealed that different siRNAs had various patterns of gene expression profiles and suggested a complicated influence of siRNAs on host cells. We further observed that 18 of these genes were suppressed by both siRNA-1 and siRNA-7. Interestingly, seven of these genes were originally activated by HBV, which suggested that these seven genes might be involved in the HBV-host cell interaction. Finally, we have compared the effects of siRNA and lamivudine on HBV and host cells, which revealed that siRNA is more effective at inhibiting HBV expression at the mRNA and protein level in vitro, and the gene expression profile of HepG2.2.15 cells treated by lamivudine is totally different from that seen with siRNA.

* Corresponding author. Mailing address: Medical Systems Biology Research Center, Tsinghua University School of Medicine, Beijing 100084, People's Republic of China. Phone: (86) 10 62772239. Fax: (86) 10 62773059. E-mail: jcheng{at}tsinghua.edu.cn.

Journal of Virology, November 2005, p. 14392-14403, Vol. 79, No. 22
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.22.14392-14403.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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