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目的 研究反义脱氧寡核苷酸 (ASON)抗乙型肝炎病毒 (HBV)复制表达的作用。方法 设计合成针对HBVPreS2 的ASON并设非互补序列作对照 ,以人肝癌细胞系 2 .2 .15细胞为细胞模型 ,应用ELISA观察了ASON对HBV基因表达的抑制作用 ,流式细胞仪观察了ASON对宿主细胞凋亡和增殖的影响。应用RIA法和MTT法观察ASON对细胞代谢的影响。结果 ASON可有效地抑制HBV基因的表达 ,对HBsAg和HBeAg的抑制率分别为 6 6 %和 91% ,而非互补序列对照无抑制作用 ;ASON可诱导宿主细胞凋亡 ,用药后第 3天和第 6天凋亡率分别为 6 .10 %和 6 .43% ,增殖指数分别为 37%和 36 % ,RIA和MTT法表明ASON对宿主细胞无毒性作用。结论 ASON不仅以序列特异性方式发挥抗病毒作用 ,而且可能以凋亡的方式清除HBV
Objective To study the effect of anti-sense oligonucleotide (ASON) on hepatitis B virus (HBV) replication and expression. Methods ASON was designed and synthesized for HBV PreS2 with non-complementary sequences as a control. The human hepatocellular carcinoma cell line 2.2.15 cells were used as a cell model. The inhibitory effect of ASON on HBV gene expression was observed by ELISA. The expression of ASON was observed by flow cytometry On host cell apoptosis and proliferation. The effect of ASON on cell metabolism was observed by RIA method and MTT method. Results ASON could effectively inhibit the expression of HBV gene and the inhibitory rates of HBsAg and HBeAg were 66% and 91%, respectively, but not non-complementary control. ASON could induce the apoptosis of host cells. On day 3 and The apoptosis rates on the 6th day were 6 .10% and 6. 43%, respectively. The proliferation index was 37% and 36%, respectively. RIA and MTT showed that ASON had no toxic effect on the host cells. Conclusion ASON not only play an antiviral effect in a sequence-specific manner, but also may clear HBV by apoptosis