Objective To construct two kinds of plasmids with one for gene expression and the other for RNA expression. Methods Hepatitis C virus (HCV) core and NS3 genes were firstly inserted into the upstream of the reporter genes marked with Rellina (hRluc) luciferase and Firefly luciferase (hluc + ). And then the established Plasmids were cotransfected into HEK293 cells. Six short hairpin RNAs targeted HCV core and NS3 genes were designed, and expressed by siRNA expression vector. Inhibition effects with the luminescent intensity were observed. Results Light density in the cells cotransfected with core gene and anti-Core siRNA was obviously weaker, meanwhile, those with NS3 gene and anti-NS3 siRNA had no difference in contrast with the control group. Transfected the anti-NS3 siRNA into the stable expression cells and detected them by real-time PCR and western-blot. The four siRNAs have efficiently suppressed the expression of NS3 at some level. Conclusion The results demonstrated that siRNA was effective in inhibiting HCV protein expression, and may have therapeutic potential to limit HCV replication in chronically infected patients. Objective To construct two kinds of plasmids with one for gene expression and the other for RNA expression. Methods Hepatitis C virus (HCV) core and NS3 genes were implanted into the upstream of the reporter genes marked with Rellina (hRluc) luciferase and Firefly luciferase (hluc +). And then the established Plasmids were cotransfected into HEK293 cells. Six short hairpin RNAs targeted HCV core and NS3 genes were designed, and expressed by siRNA expression vector. Inhibition effects with the luminescent intensity were observed. Results Light density in the cells cotransfected with core gene and anti-Core siRNA was obviously weaker, meanwhile, those with NS3 gene and anti-NS3 siRNA had no difference in contrast with the control group. Transfected the anti-NS3 siRNA into the stable expression cells and detected them by Real-time PCR and western-blot. The four siRNAs have efficiently suppressed the expression of NS3 at some level. Conclusion The results said that siRNA wa s effective in inhibiting HCV protein expression, and may have therapeutic potential to limit HCV replication in chronically infected patients.