Department of Infectious Diseases, Key Laboratory of Hunan Viral Hepatitis, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Department of Nuclear Medicine, Xiangya Hospital, Central South University, Changsha, Hunan, China.
IUBMB Life. 2019 Sep;71(9):1336-1346. doi: 10.1002/iub.2044. Epub 2019 Apr 24.
The objective of this study was to investigate the mechanism by which miR-146a-5p mediated autophagy and hepatitis B virus (HBV) replication. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the mRNA expression levels of miR-146a-5p and X-linked inhibitor of apoptosis (XIAP) and HBV DNA and RNA. The protein expression levels of XIAP, IκB-α, murine double minute 2 oncoprotein (MDM2) and p53, the phosphorylation of p65, and the conversion of light chain 3 (LC3)-I to LC3-II were detected by Western blotting. The expression levels of XIAP, HBV-related pro-inflammatory cytokines, and serum markers were detected by enzyme-linked immunosorbent assay (ELISA). miR-146a-5p was highly expressed in patients with chronic hepatitis B (CHB) and HBV-expressing hepatocytes. HBV core protein (HBc) and HBV X protein (HBx) were responsible for its effects on miR-146a-5p expression through the nuclear factor-κB pathway. Furthermore, the miR-146a-5p inhibitor suppressed autophagic response and HBV replication as well as MDM2/p53 expression. Luciferase reporter assay confirmed that XIAP was a direct target of miR-146a-5p. We therefore demonstrated that miR-146a-5p mediated positive feedback loop by regulating autophagy-induced HBV replication via targeting the XIAP-mediated MDM2/p53 axis. © 2019 IUBMB Life, 71(9):1336-1346, 2019.
本研究旨在探讨 miR-146a-5p 介导自噬和乙型肝炎病毒 (HBV) 复制的机制。采用实时定量聚合酶链反应 (qRT-PCR) 测定 miR-146a-5p 和 X 连锁凋亡抑制蛋白 (XIAP) 及 HBV DNA 和 RNA 的 mRNA 表达水平。采用 Western blot 检测 XIAP、IκB-α、鼠双微体 2 癌基因 (MDM2) 和 p53 的蛋白表达水平、p65 的磷酸化以及 LC3-I 向 LC3-II 的转化。采用酶联免疫吸附试验 (ELISA) 检测 XIAP、HBV 相关促炎细胞因子和血清标志物的表达水平。miR-146a-5p 在慢性乙型肝炎 (CHB) 患者和表达 HBV 的肝细胞中高表达。HBV 核心蛋白 (HBc) 和 HBV X 蛋白 (HBx) 通过核因子-κB 通路对 miR-146a-5p 的表达起作用。此外，miR-146a-5p 抑制剂抑制自噬反应和 HBV 复制以及 MDM2/p53 的表达。荧光素酶报告基因检测证实 XIAP 是 miR-146a-5p 的直接靶标。因此，我们证明了 miR-146a-5p 通过调节自噬诱导的 HBV 复制，通过靶向 XIAP 介导的 MDM2/p53 轴，介导正反馈环。国际生物化学与分子生物学联合会生命杂志，2019 年；71(9)：1336-1346。
