Li Qingjun, Zhang Bo, Zhang Qiling, Wang Xin, Huo Yujia, Yang Jing
Bing Du Xue Bao. 2016 Nov;32(6):747-51.
To study the effect of interferon(IFN)against hepatitis B virus(HBV)by silencing phospholipid scramblase (PLSCR)1in HepG2 cells. siRNA specific for PLSCR1 was designed and transfected in HepG2 cells. The inhibitory effect of siRNA was determined using semi-quantitative polymerase chain reaction(PCR)and western blotting 48hpost-transfection.HepG2 cells treated with IFN were co-transfected with plasmids expressing HBV1.3and siRNA targeting PLSCR1.Total RNA of HepG2 cells was isolated and the mRNA level of PLSCR1 measured by reverse-transcription semi-quantitative PCR. The expression of HBsAg in culture supernatants was determined by enzyme-linked immunosorbent assay. Expression of PLSCR1 was inhibited by siRNA911 in HepG2cells.Compared with the control, the level of HBsAg decreased in the cell supernatants of cells transfected with HBV1.3plasmid or NC-siRNA + HBV1.3plasmid.Compared with cells not treated with IFN, the level of HBsAg did not change significantly in the supernatants of cells transfected with siRNA + HBV1.3plasmid and treated with IFN. Inhibition of PLSCR1 could decrease the antiviral activity of IFN against HBV. These data suggest that PLSCR1 has an important role in the inhibition of HBV replication due to IFN.
通过沉默HepG2细胞中的磷脂翻转酶（PLSCR）1来研究干扰素（IFN）对乙型肝炎病毒（HBV）的作用。设计针对PLSCR1的小干扰RNA（siRNA）并将其转染到HepG2细胞中。转染48小时后，使用半定量聚合酶链反应（PCR）和蛋白质印迹法测定siRNA的抑制效果。用IFN处理的HepG2细胞与表达HBV1.3的质粒和靶向PLSCR1的siRNA共转染。分离HepG2细胞的总RNA，通过逆转录半定量PCR测定PLSCR1的mRNA水平。通过酶联免疫吸附测定法测定培养上清液中HBsAg的表达。siRNA911在HepG2细胞中抑制了PLSCR1的表达。与对照组相比，转染HBV1.3质粒或NC-siRNA + HBV1.3质粒的细胞的细胞上清液中HBsAg水平降低。与未用IFN处理的细胞相比，转染siRNA + HBV1.3质粒并用IFN处理的细胞的上清液中HBsAg水平没有明显变化。抑制PLSCR1可降低IFN对HBV的抗病毒活性。这些数据表明PLSCR1在IFN抑制HBV复制中起重要作用。
