关键词: 锁核酸核酶, 丙型肝炎病毒, 非编码区, 基因治疗

Abstract: Objective To investigate the inhibitory effects HCV replication of LNAzyme targeting to both 5ˊ-NCR andC genes in hepG2.9706 cells. Methods The experimental groups were divided into five groups:blank control group were treated with DMEM (dulbecco’s modified eagle medium) solution, galactose ligand control group were treated with galactose ligand alone, NCR group were treated with LNAzyme targeting to NCR gene, C group were treated with LNAzyme targeting to C gene, and dual-target group were treated with LNAzyme targeting to both NCR and C genes. LNAzyme was transfected into HepG2.9706 cells by galactose ligand. The levels of HCV RNA was quantified by Fluorescence Quantitative PCR. The expression of luciferase gene was detected by chemiluminescence technique. LNAzyme’s cyto-toxicity on cell was evaluated by MTT method. Results After LNAzyme transfection, the levels of HCV RNA in the NCR group, C group and dual-target group were reduced by 62.12%, 61.39% and 75.37%,respectively, and the luciferase gene expression were also decreased by 66.49%、65.06% and 73.30%, respectively. These values were significantly higher than those in the control groups (all P<0.05). The expression levels of fluorescent protein in the cells of all experimental groups were significantly lower than those in the control groups. Conclusion LNAzymetargeting to both 5ˊ-NCR and C gene can significant inhibit effect HCV replication and expression in vitrol, and dual-target was stronger than single target.

Key words: LNAzyme, Hepatitis C virus, Noncoding region, Gene therapy