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Objective To observe the suppression of special shRNA producing plasmid to hepatitis B virus(HBV) S gene and C gene on HBV replication and expression in HepG2.2.15 cells.Methods pSilenceCircle-U6 including pol Ⅲ promoter was used to construct HBV special shRNA producing plasmid as SC-S and SC-C.The experimental groups included SC-S group,SC-C group,unrelated control SC-N group and blank control group.With different dosages and at different time,shRNA producing plasmid was transfected into HepG2.2.15 cells.HBeAg and HBsAg in the culture media was detected by ELISA assay and HBV DNA in the culture media was measured by dot blotting assay. Results The recombinant shRNA producing plasmid with target sequence was constructed successfully.The inhibitory rates of HBeAg and HBsAg expressions by SC-S were much higher than those by SC-C.The inhibitory effects of HBeAg and HBsAg expressions were increased when the dose of SC-S was greater.The inhibitory effects of HBeAg and HBsAg expressions by SC-S were significant on the 3rd day after transfection and the inhibitory effect was the strongest on the 6th day.The inhibitory rate was still higher on the 9th day after transfection.Dot blotting assay showed the inhibitory effect of HBV replication by SC-S was greater than that by SC-C.Conclusion The shRNA producing plasmid with HBV S gene and C gene can be highly effective to inhibit the replication and expression of HBV.
RESUMEN
Objective To investigate the effect of glycyrrhizin(GL) on the expression of hepatitis B virus e antigen(HBeAg),hepatitis B virus surface antigen(HBsAg),HBV DNA levels and cell proliferation in HepG2.2.15 cell line.Methods HBV DNA level was examined by Real-time PCR.HBsAg and HBeAg levels were examined by ELASA,and cell proliferation was examined by MTT before and after stimulated with GL.The GL groups were compared with the blank control group.Results HBsAg levels in the GL groups were inhibited in dose-dependent manner compared with the blank control group(P