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目的探讨人I型干扰素受体亚基(IFNAR1)不同表达水平对IFN-α抑制HBV复制的影响。方法扩增在前期构建的IFNAR1不同表达水平稳定Hep G2细胞(R1/H1、R1/H2、R1/H3株和对照细胞PS1、PGFP细胞),分别以瞬时转染HBV表达质粒pUC19-1.24HBV和对照质粒,培养24h后加入IFN-α,48h后收集细胞采用Southern blot检测HBVD-NA,荧光定量PCR检测细胞和上清HBVDNA水平。结果 Southern blot检测提示IFN-α干预后HBV复制水平在R1/H1和R1/H2细胞及R1/H3株中IFN-α抑制细胞内核壳颗粒HBVDNA活性显著高于PS1和PGFP细胞;荧光定量PCR结果提示R1/H3细胞中细胞和上清HBVDNA水平较R1/H1、R1/H2细胞系无显著下降。结论 IFNAR1蛋白表达水平不同可能导致IFN-α抑制HBV复制效应的差异。
Objective To investigate the effect of different expression levels of human interferon type 1 receptor (IFNAR1) on the inhibition of HBV replication by IFN-α. Methods Hep G2 cells (R1 / H1, R1 / H2, R1 / H3 strain and control cells PS1 and PGFP cells) were stably transfected with HBV expression plasmids pUC19-1.24 HBV and The control plasmid was added IFN-α after culturing for 24 hours. After 48 hours, the cells were harvested for the detection of HBVD-NA by Southern blot. The level of HBVDNA in the cells and the supernatant was detected by quantitative PCR. Results The results of Southern blot showed that the level of HBV replication in IFN-α-treated cells was significantly higher than that in PS1 and PGFP cells in R1 / H1 and R1 / H2 cells and in R1 / H3 cells. Fluorescent quantitative PCR The results indicated that the level of HBVDNA in cells and supernatant of R1 / H3 cells was not significantly lower than that of R1 / H1 and R1 / H2 cell lines. Conclusion The different levels of IFNAR1 protein may result in the difference of IFN-α inhibiting HBV replication.