Identification And Verification Of Cellular Targets For Anti-HBV Drugs

Hepatitis B virus (HBV) infection is the major health concern around the world, and no effective treatment has been developed till now. The crux of the matter, limited the development of anti-HBV drugs, is due to the smaller viral genomes and the higher mutation. Several recent examples show that inhibiting host-cell proteins can prevent viral infection. The human genomes might contain more genes related to HBV infection and replication in comparing with the viral genome. A systematic approach to find these potential cellular antiviral targets is host gene expression analysis using DNA microarrays. Therefore, the aim of this study is to identify and validate the new potential cellular anti-HBV targets by using DNA microarray.Firstly, the normalization control for DNA microarray was prepared and the sensitivity of DNA microarray was detected. Then the virus-infection-related gene oligonucleiotide microarray was prepared, the results of qulity detecting indicate that this in-house microarray has high specificity, good sensitivity and stablity et al. Using this in-house microarray, the differentially expressed genes in HepG2.2.15 cells and HepG2 cells were analyzed, and also the alteration of gene expression in HepG2.2.15 cells was studied after treated by anti-HBV drugs ( lamivudine, adefovir and IBE5). Genes which were differentially expressed due to the infection of HBV but reversed by anti-HBV drugs were identified and verified by semi-quantitative RT-PCR and western blot analysis. Through bioinformatics analysis, ASGPR1 and fibronectin were speculated to have the possibility of being the new potential cellular anti-HBV targets, that is, ASGPR1 or fibronectin inhibition might block HBV replication in HepG2.2.15 cells. To support our speculations, antisense oligodeoxynucleotides, ligands, antibodies and other methods were used to inhibit the expression of potential targets. The results strongly suggest that ASGPR1 or fibronectin inhibition dose-dependently block HBV replication in HepG2.2.15 cells, which provide strong evidence for the possibility of...