针对HBV调节基因ENⅡ的ASON抑制HBV复制表达的体外研究

In vitro Study on Inhibition of HBV Replication and Expression by Antisense Oligodeoxynucleotides toward HBV Regulation Gene ENⅡ

目的 :筛选出高效特异而无毒的抗HBV反义寡核苷酸片段。方法 :以 2 2 15细胞为模型 ,设计合成针对HBVENⅡ的反义硫代寡苷酸 (ASON) ,ELISA法检测ASON对宿主细胞分泌表达HBsAg和HBeAg的抑制作用 ,MTT法检测ASON对细胞增殖代谢的影响。结果 :ASON对HBsAg和HBeAg的抑制率分别为 92 %和 75 % ,与非互补序列对照组 (抑制率均为 11% )相比差异有显著性意义 (P <0 0 0 1) ,用药前后MTT比色结果和活细胞比率差异无显著性意义 (P >0 0 5 )。结论

Objective:To screen the high performance,specific and nontoxic anti HBV antisense oligonucleotide fragment.Methods:Taking 2.2.15 cells as cell model,the antisense oligodeoxynucleotides(ASON)toward the HBV regulation genesenhancerⅡ(HBV EFⅡ)were designed and synthesized.The role played by ASON in inhibiting the secretion and expession of HBsAg and HBeAg by the host cells was detected by ELISA method.The effect of ASON on prolifezation and metabolism of the cells was detected by MTT method.Results:The inhibitory rate of HBsAg by ASON was 92% and 75%,respectively,indicating that itsdifference from that of noncomplementary sequence control group(inhibitory rate was 11%)had considerable statistical significance( P <0.001).The difference between MTT colorimetric result and living cell ratio prior and post drug administration had no considerable statistical significunce( P >0.05).Conclusion:ENⅡ is one of the most important target oriented sequencial selective regions of research on anti HBV nature of antisense oligonucleotide.