Searching for cellular factors facilitating HCV replication

• Main Authors: Wan-Ling Hsiao, 蕭婉玲
• Other Authors: Shih-Yen Lo
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/55177844441879113708

碩士 === 慈濟大學 === 醫學生物技術研究所 === 96 === Hepatitis C virus ( HCV ) infects more than 170 million people worldwide, leading to both acute and chronic liver diseases in patients. Hepatitis C virus ( HCV ) is an enveloped virus with a single stranded 9.6-kb RNA genome. It is a member of the Flaviviridae. Nowadays, the combined therapy with interferon-alpha ( IFN-α ) and ribavirin ( RBV ) remains the only available option for treatment of patients with chronic hepatitis C. There is no vaccine available and the standard therapy [ ( pegylated ) interferon alfa plus ribavirin ] is only effective in 50–60% of patients and is associated with important side-effects. Cyclosporin A ( CsA ) , a widely used immunosuppressive drug, has been reported to be effective against HCV infection. The antiviral action of CsA is mediated by blockade of actions of cellular CsA-binding proteins, the cyclophilins. Therefore, cellular cofactors affecting hepatitis C virus replication could be the potential anti-viral targets. In this study, we are going to search for the cellular factors that can facilitate HCV replication. Microarray analysis was used to screen the genes with differential expression level between the HCV replicon cell line and mutant cell line without HCV replicon. Some of these differentially expressed genes were further verified by RT-qPCR. Genes over-expressed in HCV replicon were knocked down by shRNA technology individually to determine their effect on the HCV replication. The replication of HCV was monitored by the expression of HCV NS5A protein. The cellular factor KCNJ8 was identified. Then, we used its inhibitor PNU-37883A to block its expression and detected the expression of HCV NS5A protein. We found that KCNJ8 could affect HCV replication. Moreover, the combination treatment of either interferon-α and PNU-37883A or CsA and PNU-37883A could inhibit HCV replication more effectively than that of interferon-α or CsA treatment alone. These results indicate that KCNJ8 could be a potential antiviral target to limit HCV replication.