Role of ISGF3 in modulating the anti-hepatitis B virus activity of interferon-alpha in vitro
Role of ISGF3 in modulating the anti-hepatitis B virus activity of interferon-alpha in vitro
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ISGF3在体外调节干扰素-α抗乙型肝炎病毒活性中的作用
DOI：
10.1111/j.1440-1746.2007.04985.x
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发表时间：
2008-11-01
影响因子：
4.1
通讯作者：
Cong, Xu
中科院分区：
文献类型：
--
作者：
Zhang, Quan;Wang, Yan;Cong, Xu
尽管干扰素-α（IFN-α）是治疗乙型肝炎病毒（HBV）感染的有效方法，但其确切的作用机制尚未确定。在本研究中，我们研究了信号转导通路在 IFN-α 介导的抗 HBV 反应激活中的作用。使用寡核苷酸微阵列，我们发现人肝癌细胞中 IFN-α 显着上调 IFN-α 信号通路中的四个基因，无论它们是否转染了含有 HBV 基因组的质粒：信号转导子和激活子 转录 1 (STAT1)、干扰素调节因子 9 (IRF-9，也称为 ISGF3 gamma 或 P48)、IFN-α 诱导蛋白 15 (IFI-15) 和 IFN-α 诱导蛋白 6-16 (IFI-6-16)。我们还通过使用半定量逆转录 PCR (RT-PCR) 测量 ISGF3 内的三个基因（STAT1、STAT2 和 IRF-9）的 mRNA，并通过蛋白质印迹法测量这三种蛋白的表达，以及 dsRNA 依赖性蛋白激酶 (PKR)。IFN-α 治疗可上调 STAT1、STAT2、IRF-9 和 PKR mRNA 以及蛋白水平。当细胞用金雀异黄素预处理时，IFN-α刺激后STAT1、STAT2和IRF-9 mRNA水平保持不变，但PKR mRNA水平下降，STAT1、P-STAT2、IRF-9和PKR蛋白表达下降。用 IFN-α 处理的细胞上清液中 HBV DNA 水平降低，而 ISGF3 水平升高。用金雀异黄素预处理后，HBV DNA 的数量保持不变。这些观察结果表明，Janus 酪氨酸激酶-STAT (JAK-STAT) 通路可能在介导 IFN-α 对抗 HBV 的作用中发挥主要作用，而 ISGF3 可能是一个关键因素。
Although interferon-alpha (IFN-alpha) is an effective treatment for hepatitis B virus (HBV) infection, its precise mechanism of action has not been identified. In this study, we investigated the role of signal transduction pathways in the activation of anti-HBV responses mediated by IFN-alpha.Using an oligo microarray, we found that four genes in the IFN-alpha signal pathway were markedly upregulated by IFN-alpha in human hepatoma cells regardless of whether they had been transfected with a plasmid containing the HBV genome: signal transducers and activators of transcription 1 (STAT1), interferon regulatory factor-9 (IRF-9, also called ISGF3 gamma or P48), IFN-alpha-inducible protein 15 (IFI-15) and IFN-alpha-inducible protein 6-16 (IFI-6-16). We also investigated the role of IFN-stimulated gene factor3 (ISGF3) complex in IFN-alpha-mediated anti-HBV responses in human hepatoma cells by measuring the mRNA of the three genes within ISGF3 (STAT1, STAT2 and IRF-9) using semiquantitative reverse-transcription PCR (RT-PCR), and expression of the three proteins by western blot, and the mRNA and protein of dsRNA-dependent protein kinase (PKR).STAT1, STAT2, IRF-9 and PKR mRNA as well as protein levels were upregulated by IFN-alpha treatment. When cells were pretreated with genistein, STAT1, STAT2 and IRF-9 mRNA levels remained unchanged after IFN-alpha stimulation, but PKR mRNA levels decreased, and the expression of the STAT1, P-STAT2, IRF-9 and PKR proteins decreased. Levels of HBV DNA decreased in the supernatants of cells treated with IFN-alpha, while ISGF3 levels increased. The quantity of HBV DNA remained unchanged by pretreating with genistein.These observations suggested that the Janus tyrosine kinase-STAT (JAK-STAT) pathway may play a major role in mediating the effects of IFN-alpha against HBV, and that ISGF3 might be a key factor.