Downregulation of autophagy-related gene ATG5 and GABARAP expression by IFN-λ1 contributes to its anti-HCV activity in human hepatoma cells
Downregulation of autophagy-related gene ATG5 and GABARAP expression by IFN-λ1 contributes to its anti-HCV activity in human hepatoma cells
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DOI：
10.1016/j.antiviral.2017.01.016
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发表时间：
2017-04-01
影响因子：
7.6
通讯作者：
Liang, Hao
中科院分区：
文献类型：
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作者：
Li, Xu;Li, Yu;Liang, Hao
III型干扰素（IFN-λ）是最近发现的IFN家族，与I型IFN具有共同的特征，但也具有许多独特的活性。目前尚不清楚IFN-λ是否具有额外的抗病毒机制。在这项研究中，我们研究了IFN-λ对自噬的影响，自噬是一种与丙型肝炎病毒（HCV）感染人肝癌Huh 7细胞密切相关的细胞过程。我们的结果显示IFN-λ 1处理抑制Huh 7细胞中的自噬活性，如通过微管相关蛋白1轻链3B（LC 3B）-II的表达降低和LOB-I向LOB-II的转化、GFP-LO斑点的形成减少和自噬体的积累减少所证明的。IFN-λ I还可以抑制HCV诱导的或衣霉素（一种已知的自噬诱导剂，具有与HCV感染相似的机制）诱导的LOB-II表达和自噬体形成。通过PCR芯片、真实的时间RT PCR和western blot，鉴定了两个与自噬相关的基因ATG 5和GABARAP，并证实IFN-λ 1在未感染HCV的Huh 7细胞和感染HCV的JFH-1细胞中下调。过表达ATG 5和/或GABARAP可部分恢复IFN-λ 1抑制的HCV复制。机制研究表明，IFN-A1可诱导miR-181 a和miR-214（分别靶向ATG 5和GABARAP）的表达，从而下调ATG 5和GABARAP的表达。总之，我们的结果表明，IFN-λ 1抑制自噬反应有助于IFN-λ 1抗HCV活性。该结果也为通过抑制HCV诱导的自噬反应来提高IFN治疗HCV感染的有效性提供了理论依据。
Type-Ill interferon (IFN-lambda), the most recently discovered family of IFNs, shares common features with type I IFNs, but also has many distinctive activities. It is not clear that whether IFN-lambda has additional antiviral mechanisms. In this study, we investigated the effects of IFN-lambda on autophagy, a cellular process closely related to hepatitis C virus (HCV) infection in human hepatoma Huh7 cells. Our results showed that IFN-lambda 1 treatment inhibit autophagic activity in Huh7 cells, as evidenced by the decreased expression of microtubule-associated protein 1 light chain 3B (LC3B)-II and conversion of LOB-I to LOB -II, decreased formation of GFP-LO puncta and accumulation of autophagosomes. IFN-lambda I could also inhibit HCV-induced or tunicamycin (a known inducer of autophagy with similar mechanism to HCV infection)-induced LOB -II expression and autophagosome formation. Through PCR array, real time RT PCR, and western blot, two autophagy-related genes, ATG5 and GABARAP, were identified and verified to be down-regulated by IFN-lambda 1 treatment, either in HCV-uninfected Huh7 cells or in HCV JFH-1-infected cells. Overexpression of ATG5 and/or GABARAP could partly recover the IFN-lambda 1-inhibited HCV replication. Mechanism research demonstrated that IFN-A1 could induce the expression of miR-181a and miR-214 (targeting ATG5 and GABARAP respectively), by which down-regulates ATG5 and GABARAP expression. Taken together, our results indicate that suppression of the autophagy response by IFN-lambda 1 contributes to IFN-lambda 1 anti-HCV activity. The results also provide a theoretical basis for improving the effectiveness of IFN treatment of HCV infection through inhibition of the HCV-induced autophagy response.