不同方式给予反义寡核苷酸对培养肝癌细胞HBV基因表达的影响

Inhibitory effect of antisense oligodeoxynucleotide on HBV gene expression given by different ways

为开发新型抗HBV药物 ,设计合成针对HBVENⅡ (1713 172 7)的反义寡核苷酸 (as ENⅡ ) ,以不同浓度、不同作用方式作用于HepG2 2 15细胞 ,ELISA检测结果表明 10 μmol/L的as ENⅡ可显著抑制HBsAg、HBeAg表达 ,抑制率可达 5 2 1%和 40 1%。 10 μmol/Las ENⅡ一次性用于HepG2 2 15细胞后 ,其对HBsAg、HBeAg的抑制作用呈现双峰现象 :分别在 1d、5d和 1d、3d ,抑制作用分别为 30 4% ,5 2 1%和 34 0 % ,40 1%。同样浓度as ENⅡ每天反复加入HepG2 2 15细胞 ,其抑制作用缓慢上升 ,分别在第 7、6d达高峰 ,抑制作用分别为 73 6 9%和 73 6 4%。结果提示HBVENⅡ是设计抗HBV反义寡核苷酸片段的良好靶位 。

To explore new therapy against HBV, the inhibitory effect on HBV gene expression of antisense oligodeoxynucleotide targeting to enhancer Ⅱ of HBV (as ENⅡ) was assayed by ELISA Results showed that 10μmol/L of as ENⅡ inhibited HBV gene expression significantly The inhibitory rates of HBsAg and HBeAg were 52 1% and 40 1% respectively The inhibitory effect on expression of these antigens was bimodel when 10μmol/L as ENⅡ was given in one time It was found that the two inhibitory peaks on HBsAg expression appeared on the 1st and 5th day after as ENⅡ was added to HepG2 2 15 cells(30 3%,50 2%),while the two inhibitory peaks on HBeAg appeared on the 1st and 3rd day (34%,40%) When the same ammount of as ENⅡ was given every day within 7 days,the inhibitory effect on HBsAg and HBeAg rose gradually ,then reached its highest inhibition on the 7th and 6th day respectively(73 69%,73 34%) This study may have important implications for the therapeutic use of antisense ologodeoxynucleotide in the context of HBV infection