摘要
AIM:To observe the inhibition of hepatitis B virus replication and expression by combinated siRNA and lamivudine in HepG2.2.15 cells.METHODS:Recombinant plasmid psil-HBV was constructed and transfected into HepG2.2.15 cells.The transfected cells were cultured in lamivudine containing medium(0.05um) and harvested at 48,72 and 96 h.The concentration of HBeAg and HBsAg was determined using ELISA method.HBV DNA replication was examined by real time PCR and the level of HBV mRNA was measured by reserved transcribed PCR.RESULTS:In HepG2.2.15 cells treated with combinated siRNA and lamivudine,the secretion of HBeAg and HBsAg into the supernatant was found to be inhibited by 91.80%and 82.40%(P < 0.05) at 96 h,the number of HBV DNA copies within culture supernatants was also significantly decreased at 96 h(P < 0.05);the concentration of mRNA was lower 80.56%than the single use either siRNA or lamivudine(P < 0.05).CONCLUSION:Combination of siRNA and lamivudine is more effective in inhibiting HBV replication than the use of any one in HepG2.2.15 cells.
AIM:To observe the inhibition of hepatitis B virus replication and expression by combinated siRNA and lamivudine in HepG2.2.15 cells.METHODS:Recombinant plasmid psil-HBV was constructed and transfected into HepG2.2.15 cells.The transfected cells were cultured in lamivudine containing medium(0.05um) and harvested at 48,72 and 96 h.The concentration of HBeAg and HBsAg was determined using ELISA method.HBV DNA replication was examined by real time PCR and the level of HBV mRNA was measured by reserved transcribed PCR.RESULTS:In HepG2.2.15 cells treated with combinated siRNA and lamivudine,the secretion of HBeAg and HBsAg into the supernatant was found to be inhibited by 91.80%and 82.40%(P < 0.05) at 96 h,the number of HBV DNA copies within culture supernatants was also significantly decreased at 96 h(P < 0.05);the concentration of mRNA was lower 80.56%than the single use either siRNA or lamivudine(P < 0.05).CONCLUSION:Combination of siRNA and lamivudine is more effective in inhibiting HBV replication than the use of any one in HepG2.2.15 cells.
引文