关键词: 脂质体, 乙型肝炎病毒, 锁核酸, 反基因治疗

Abstract: Objective To investigate the inhibitory effects of hepatitis B virus(HBV) preS1 gene-specific anti-gene locked nucleic acid(LNA) on HBV replication and expression in HepG2.2.15 cells. Methods The anti-gene LNA which were complementary to the purine rich region of HBV preS1 gene were designed, synthesized, and transfected by cationic liposomes into HepG2 2.2.15 cells. The HBsAg,preS1-Ag and HBV DNA of supernatant was tested by time-resolved fluorescence immune assay(TRFIA) ,real-time fluorescent quantitative PCR(FQ-PCR) and enzyme linked immunosorbent assays(ELISA) at 1, 3, 5 and 7 d after treatment. LNA’s cyto-toxicity on cell was evaluated by MTT method. Results The anti-gene LNA that targeting on the purine rich region of HBV preS1 gene showed strong inhibitory effects on replication of HBV DNA and the expression of HBsAg and preS1-Ag with the inhibition rates of 65.99%,67.49% and 63.88% respectively after 7 days.There’s no obvious toxicity on cell. Conclusion anti-gene locked nucleic acid(LNA) that targeting on the purine rich region of HBV preS1 gene has show strong inhibition on HBV in vitro.It has a therapeutic potential in the treatment of patients infected with HBV.

Key words: Cationic liposomes, Hepatitis B virus, Locked nucleic acid, anti-gene therapy