Hepatitis B virus-specific miRNAs and Argonaute2 play a role in the viral life cycle.
Hepatitis B virus-specific miRNAs and Argonaute2 play a role in the viral life cycle.
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DOI：
10.1371/journal.pone.0047490
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Chayama K
中科院分区：
文献类型：
--
作者：
Hayes CN;Akamatsu S;Tsuge M;Miki D;Akiyama R;Abe H;Ochi H;Hiraga N;Imamura M;Takahashi S;Aikata H;Kawaoka T;Kawakami Y;Ohishi W;Chayama K
疾病特异性血清miRNA谱可作为生物标志物，并可能揭示潜在的新治疗途径。最近报道了与HBV表面抗原（HBsAg）颗粒相关的HBV特异性血清miRNA谱，并且AGO 2和miRNA已显示与血清中的HBsAg稳定相关。我们使用Toray 3D阵列系统在10名健康对照和10名慢性肝炎B病毒（HBV）感染患者中鉴定HBV相关血清miRNAs。然后通过定量RT-PCR检测248例慢性HBV患者和22例健康对照者的19种miRNAs。还使用活检样品比较了血清与肝组织中的miRNA表达。为了研究AGO 2在HBV生命周期中的作用，我们在稳定转染的HepG 2细胞中使用免疫细胞化学和邻位连接测定法分析了AGO 2和HBV核心（HBcAg）和表面（HBsAg）抗原的细胞内共定位。使用siRNA评估AGO 2消融对病毒复制的影响。在HBV感染患者血清中，包括miR-122、miR-22和miR-99 a在内的几种miRNA上调至少1.5倍（P<2 E-08）。AGO 2和HBcAg被发现在ER和其他亚细胞区室中物理相互作用和共定位。还发现HBs与AGO 2共定位，并在多个亚细胞区室中检测到。相反，HBx非特异性地定位于细胞核和细胞质中，并且未检测到AGO 2和HBx之间的相互作用。siRNA阻断AGO 2可抑制上清液中HBV DNA和HBs抗原的产生。这些结果表明，AGO 2和HBV特异性miRNA可能在HBV生命周期中发挥作用。
Disease-specific serum miRNA profiles may serve as biomarkers and might reveal potential new avenues for therapy. An HBV-specific serum miRNA profile associated with HBV surface antigen (HBsAg) particles has recently been reported, and AGO2 and miRNAs have been shown to be stably associated with HBsAg in serum. We identified HBV-associated serum miRNAs using the Toray 3D array system in 10 healthy controls and 10 patients with chronic hepatitis B virus (HBV) infection. 19 selected miRNAs were then measured by quantitative RT-PCR in 248 chronic HBV patients and 22 healthy controls. MiRNA expression in serum versus liver tissue was also compared using biopsy samples. To examine the role of AGO2 during the HBV life cycle, we analyzed intracellular co-localization of AGO2 and HBV core (HBcAg) and surface (HBsAg) antigens using immunocytochemistry and proximity ligation assays in stably transfected HepG2 cells. The effect of AGO2 ablation on viral replication was assessed using siRNA. Several miRNAs, including miR-122, miR-22, and miR-99a, were up-regulated at least 1.5 fold (P<2E-08) in serum of HBV-infected patients. AGO2 and HBcAg were found to physically interact and co-localize in the ER and other subcellular compartments. HBs was also found to co-localize with AGO2 and was detected in multiple subcellular compartments. Conversely, HBx localized non-specifically in the nucleus and cytoplasm, and no interaction between AGO2 and HBx was detected. SiRNA ablation of AGO2 suppressed production of HBV DNA and HBs antigen in the supernatant. These results suggest that AGO2 and HBV-specific miRNAs might play a role in the HBV life cycle.
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