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研究磷脂爬行酶1(Phospholipid scramblase 1,PLSCR1)对干扰素抑制HBV作用的影响。设计合成PLSCR1特异性小干扰RNA(siRNA),以完全随机序列的阴性小干扰(NCsiRNA)作为对照,转染HepG2细胞,于转染48h后分别检测PLSCR1mRNA和蛋白水平表达量的变化,筛选出对PLSCR1具有沉默作用的siRNA;将HepG2细胞分为正常对照组和干扰素处理组,将1.3倍乙型肝炎病毒(HBV)全基因真核细胞表达载体HBV1.3质粒分别与PLSCR1siRNA或NCsiRNA共同转染HepG2细胞或干扰素处理的HepG2细胞,转染48h后检测各组细胞中PLSCR1mRNA表达量及培养液上清中HBsAg表达水平。PLSCR1特异性小干扰RNA siRNA911转染后能够显著抑制HepG2细胞中PLSCR1基因在mRNA和蛋白水平的表达;与HepG2细胞对照组比较,干扰素处理组细胞转染HBV1.3质粒、NCsiRNA+HBV1.3质粒后,细胞培养液中HBsAg表达水平均显著降低(P<0.05);而PLSCR1siRNA与HBV1.3共转染IFN处理的HepG2细胞组与共转染HepG2细胞组相比较,细胞培养液中HBsAg的表达水平没有显著差异。提示抑制PLSCR1的siRNA可抑制干扰素的抗HBV活性,提示PLSCR1在干扰素抑制HBV复制中具有重要作用。
To investigate the effect of Phospholipid scramblase 1 (PLSCR1) on the inhibition of HBV by interferon. The specific small interfering RNA (siRNA) of PLSCR1 was designed and synthesized. HepG2 cells were transfected with completely random sequences of negative small interfering RNA (NCsiRNA), and the expression of PLSCR1 mRNA and protein were detected 48 h after transfection. PLSCR1 silenced siRNA; HepG2 cells were divided into normal control group and interferon treatment group, 1.3 times of hepatitis B virus (HBV) whole genome eukaryotic cell expression vector HBV1.3 plasmid were co-transfected with PLSCR1 siRNA or NCsiRNA HepG2 cells or HepG2 cells treated with interferon. The expression of PLSCR1 mRNA and the level of HBsAg in the culture supernatants were detected 48 h after transfection. The expression of PLSCR1 mRNA and protein in HepG2 cells was significantly inhibited by PLSCR1 siRNA siRNA1111 transfection. Compared with HepG2 cells control group, the interferon treated cells transfected with HBV1.3 plasmid, NCsiRNA + HBV1.3 Compared with HepG2 cells transfected with IFN-treated HepG2 cells transfected with PLSCR1 siRNA and HBV1.3, HepG2 cells transfected with HepG2 cells transfected with PLSCR1 siRNA showed that the expression of HBsAg in cell culture medium was significantly lower than that in HepG2 cells transfected with plasmids (P <0.05) There are no significant differences in levels. It is suggested that siRNAs that inhibit PLSCR1 can inhibit the anti-HBV activity of interferon, suggesting that PLSCR1 plays an important role in the inhibition of HBV replication by interferon.