环状RNA circBACH1通过调控miR-200a-3p/MAP3K2轴促进乙型肝炎病毒复制和肝癌发展。
CircRNA circBACH1 facilitates hepatitis B virus replication and hepatoma development by regulating the miR-200a-3p/MAP3K2 axis.
作者信息
Du Na, Li Kailin, Wang Yu, Song Bo, Zhou Xuan, Duan Shaoqiong
机构信息
Department of Infectious Disease, Suizhou Central Hospital Affiliated to Hubei Medical College, Suizhou, China.
Department of Cardiology, Suizhou Central Hospital Affiliated to Hubei Medical College, Suizhou, China.
出版信息
Histol Histopathol. 2022 Sep;37(9):863-877. doi: 10.14670/HH-18-452. Epub 2022 Feb 3.
BACKGROUND
Hepatitis B virus (HBV) is a top contributor to hepatoma. Circular RNAs (circRNAs) have been elucidated to have a close connection with HBV-induced hepatoma. This study aimed to explore the role of circRNA BTB domain and CNC homolog 1 (circBACH1) in HBV replication and hepatoma progression, as well as the potential mechanistic pathway.
METHODS
Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed to assess the expression of circBACH1, microRNA (miR)-200a-3p, and mitogen-activated protein kinase kinase kinase 2 (MAP3K2). HBV replication was determined by enzyme-linked immunosorbent assay (ELISA) and qRT-PCR assay. Cell viability and clonogenicity were detected via Cell Counting Kit-8 (CCK-8) assay and colony formation assay, respectively. Cell metastasis was examined by Transwell assay and wound healing assay. Annexing-V/PI staining was employed to monitor cell apoptosis using flow cytometry. Levels of MAP3K2, proliferation- and apoptosis-related proteins were analyzed by Western blotting. Target interaction between miR-200a-3p and circBACH1 or MAP3K2 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The role of circBACH1 in vivo was investigated by xenograft model assay.
RESULTS
Expression of circBACH1 and MAP3K2 was increased, while miR-200a-3p expression was decreased in HCC tissues and HBV-transfected hepatoma cells. Depletion of circBACH1 or miR-200a-3p overexpression impeded HBV replication, proliferation, and metastasis in HBV-transfected hepatoma cells. CircBACH1 was able to regulate MAP3K2 expression by sponging miR-200a-3p. CircBACH1 regulated HBV replication and hepatoma progression through the miR-200a-3p/MAP3K2 pathway. Moreover, circBACH1 deficiency hampered tumor growth in vivo.
CONCLUSION
CircBACH1 knockdown had inhibitory effects on HBV replication and hepatoma progression, at least partly by modulating the miR-200a-3p/MAP3K2 axis.
背景
乙型肝炎病毒（HBV）是肝癌的主要致病因素之一。环状RNA（circRNAs）已被证实与HBV诱导的肝癌密切相关。本研究旨在探讨环状RNA BTB结构域和CNC同源蛋白1（circBACH1）在HBV复制和肝癌进展中的作用及其潜在机制。
方法
采用定量实时聚合酶链反应（qRT-PCR）检测circBACH1、微小RNA（miR）-200a-3p和丝裂原活化蛋白激酶激酶激酶2（MAP3K2）的表达。通过酶联免疫吸附测定（ELISA）和qRT-PCR检测HBV复制情况。分别采用细胞计数试剂盒-8（CCK-8）法和集落形成试验检测细胞活力和克隆形成能力。通过Transwell试验和伤口愈合试验检测细胞迁移情况。采用Annexin-V/PI染色，通过流式细胞术监测细胞凋亡。通过蛋白质免疫印迹法分析MAP3K2、增殖和凋亡相关蛋白的水平。通过双荧光素酶报告基因试验和RNA免疫沉淀（RIP）试验证实miR-200a-3p与circBACH1或MAP3K2之间的靶向相互作用。通过异种移植模型试验研究circBACH1在体内的作用。
结果
在肝癌组织和HBV转染的肝癌细胞中，circBACH1和MAP3K2的表达升高，而miR-200a-3p的表达降低。circBACH1的缺失或miR-200a-3p的过表达可抑制HBV转染的肝癌细胞中的HBV复制、增殖和迁移。CircBACH1能够通过吸附miR-200a-3p来调节MAP3K2的表达。CircBACH1通过miR-200a-3p/MAP3K2途径调节HBV复制和肝癌进展。此外，circBACH1的缺失阻碍了体内肿瘤的生长。
结论
CircBACH1的敲低对HBV复制和肝癌进展具有抑制作用，至少部分是通过调节miR-200a-3p/MAP3K2轴实现的。
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