Objective: To study the effect of HCV NS5A protein on HCV IRES-dependent translation in HepG2 cells.
Methods: HepG2 cells were co-transfected with a plasmid vector containing a bicistronic transcript carrying Renilla luciferase and firefly luciferase genes separated by HCV IRES sequences, and an expressing vector producing the NS5A protein. The luciferase activity and the mRNA of the luciferase gene were then detected. The NS5A expression was confirmed by fluorescence microscopy.
Results: HCV NS5A protein was detected in the cytoplasm of the HepG2 cells transfected with pcDNA-NS5A, and the luciferase activity was up-regulated in the presence of the HCV NS5A protein while the expression of luciferase mRNA showed no difference.
Conclusion: HCV NS5A protein can upregulate the HCV IRES activity and this effect is dose-dependent with NS5A.