硫代磷酸反义寡核苷酸对乙型肝炎病毒抗原表达的体外抑制作用

ANTISENSE PHOSPHOROTHIOATE OLIGONUCLEOTIDES INHIBITION THE EXPRESSION OF GENE FOR HEPATITIS B VIRUS ANTIGENS IN VITRO

目的研究硫代磷酸反义寡核苷酸(S-ASON)的抗乙型肝炎病毒(HBV)作用.方法作者以2.2.15细胞作为研究对象,在 HBV 基因 S 区和 C 区的翻译起始位点设计合成了2段16聚 S-ASONs,采用酶联免疫吸附试验检测了培养细胞上清中表面抗原(HBsAg)和 e 抗原(HBcAg)的分泌情况。结果当 S-ASON 浓度为每天2μmol/L 时,对 HBsAg 和HBcAg 的抑制率可分别达到88%和75%;而无关序列的寡核苷酸对 HBsAg 和 HBeAg 的抑制作用甚小(抑制率≤15%);ASONs 未对细胞产生毒性作用;ASONs 对 HBV 基因表达的序列特异性抑制作用呈剂量依赖性方式.结论 S-ASON可能成为抗 HBV 的一种有效制剂.

Objective In order to study the action of antisense phosphorothioate oligonucleotides(S-ASON)to hepatitis B virus(HBV).Methods The 2.2.15 cells were chosen as an in vitro cell culture assay system and S-ASONs were synthesized against the translation initiation sites of the S gene and the C gene of HBV.Hepatitis B surface antigen (HBsAg)and hepatitis B e antigen(HBeAg)in cell culture medium were tested by enzyme linked immunosorbent assay.Results S-ASONs could inhibit 88% of the HBsAg and 75% of the HBeAg produced at a concentration of 2 μmol/L per day.S-ASON had no cytotoxicity.There was minimal(≤15%),if any reduction of HBsAg and HBeAg by the cells treated with similar concentration of an unrelated sequence oligonucleotide.S-ASONs showed a does- dependent and sequence-specific inhibition of HBV gene expression.Conclusion These results suggest a therapeutic potential effect for S-ASON on treatment for patients with HBV.