title: Genomic analysis of anti-hepatitis B virus (HBV) activity by small interfering RNA and lamivudine in stable HBV-producing cells.
abstract: 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.
authors.affiliation: Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Guo
authors.forename: Hongyan
authors.initials: H
authors.lastname: Guo
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.forename: Hongying
authors.initials: H
authors.lastname: Xie
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Fangxun
authors.initials: F
authors.lastname: Wang
authors.forename: Ze
authors.initials: Z
authors.lastname: Li
authors.forename: Yahui
authors.initials: Y
authors.lastname: Wang
authors.forename: Shiliang
authors.initials: S
authors.lastname: Ma
authors.forename: Jiaping
authors.initials: J
authors.lastname: Tao
authors.forename: Weixing
authors.initials: W
authors.lastname: Wang
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Zhou
authors.forename: Weiping
authors.initials: W
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Cheng
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D005822:Genetic Vectors; D016679:Genome, Viral; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology*; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D012367:RNA, Viral / Q000187:drug effects / Q000235:genetics; D015203:Reproducibility of Results; D015183:Restriction Mapping; D014162:Transfection
chemical_list: D000998:Antiviral Agents; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D019259:Lamivudine