A positive-feedback loop between HBx and ALKBH5 promotes hepatocellular carcinogenesis.
A positive-feedback loop between HBx and ALKBH5 promotes hepatocellular carcinogenesis.
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DOI：
10.1186/s12885-021-08449-5
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发表时间：
2021-06-10
期刊：
影响因子：
3.8
通讯作者：
Zeng Z
中科院分区：
文献类型：
--
作者：
Qu S;Jin L;Huang H;Lin J;Gao W;Zeng Z
B型肝炎病毒（HBV）通过多种表观遗传机制参与肝癌的发生。新近定义的表观遗传学，即表观转录组学调控，已被报道涉及包括肝细胞癌（HCC）在内的多种癌症。我们前期的研究发现HBx（HBV编码X蛋白）以WDR 5依赖的方式介导H3 K4 me 3修饰参与HBV感染并促进癌基因表达。AlkB同系物5（ALKBH 5）是一种与多种肿瘤相关的表位转录酶。然而，ALKBH 5是否以及如何在HBV相关的HCC中失调仍不清楚。本研究旨在探讨ALKBH 5在中国人HBV相关性肝癌（HBV-HCC）患者中的功能、临床意义及作用机制。通过RT-qPCR、Western blot、数据挖掘和免疫组织化学方法评估ALKBH 5在总共373个HBV-HCC组织和4个HCC细胞系中的表达模式。进行细胞计数试剂盒8（CCK 8）测定、Transwell和裸鼠模型以评估小干扰RNA和慢病毒颗粒两者对ALKBH 5的功能。通过CHIP-qPCR在HBx和WDR 5敲低细胞中确定ALKBH 5的调节机制。在HBV驱动的细胞和HBx过表达细胞中，通过MeRIP-qPCR和放线菌素D抑制剂实验进一步评价ALKBH 5在HBx mRNA N6-甲基腺苷（m6 A）修饰中的作用。ALKBH 5在肿瘤组织中增加，并预测HBV-HCC的预后不良。在机制上，在HBV感染后，通过HBx介导的H3 K4 me 3修饰ALKBH 5基因启动子以WDR 5依赖性方式诱导高度表达的ALKBH 5。增加的ALKBH 5蛋白催化HBx mRNA的m6 A去甲基化，从而稳定并有利于更高的HBx表达水平。HBx和ALKBH 5在HBV-HCC组织中存在正相关性，ALKBH 5的缺失在体内外均能显著抑制HBV驱动的肿瘤细胞的生长和迁移。HBx-ALKBH 5可能形成一个正反馈环参与HBV诱导的肝癌发生，以ALKBH 5为靶点治疗HBV-HCC可能是一条潜在的途径。在线版本包含补充材料，可通过10.1186/s12885-021-08449-5获得。
Hepatitis B Virus (HBV) contributes to liver carcinogenesis via various epigenetic mechanisms. The newly defined epigenetics, epitranscriptomics regulation, has been reported to involve in multiple cancers including Hepatocellular Carcinoma (HCC). Our previous study found that HBx, HBV encodes X protein, mediated H3K4me3 modification in WDR5-dependent manner to involve in HBV infection and contribute to oncogene expression. AlkB Homolog 5 (ALKBH5), one of epitranscriptomics enzymes, has been identified to be associated with various cancers. However, whether and how ALKBH5 is dysregulated in HBV-related HCC remains unclear yet. This study aims to investigate ALKBH5 function, clinical significance and mechanism in HBV related HCC (HBV-HCC) patients derived from Chinese people. The expression pattern of ALKBH5 was evaluated by RT-qPCR, Western blot, data mining and immunohistochemistry in total of 373 HBV-HCC tissues and four HCC cell lines. Cell Counting Kit 8 (CCK8) assay, Transwell and nude mouse model were performed to assess ALKBH5 function by both small interference RNAs and lentiviral particles. The regulation mechanism of ALKBH5 was determined in HBx and WDR5 knockdown cells by CHIP-qPCR. The role of ALKBH5 in HBx mRNA N6-methyladenosine (m6A) modification was further evaluated by MeRIP-qPCR and Actinomycin D inhibitor experiment in HBV-driven cells and HBx overexpression cells. ALKBH5 increased in tumor tissues and predicts a poor prognosis of HBV-HCC. Mechanically, the highly expressed ALKBH5 is induced by HBx-mediated H3K4me3 modification of ALKBH5 gene promoter in a WDR5-dependent manner after HBV infection. The increased ALKBH5 protein catalyzes the m6A demethylation of HBx mRNA, thus stabilizing and favoring a higher HBx expression level. Furthermore, there are positive correlations between HBx and ALKBH5 in HBV-HCC tissues, and depletion of ALKBH5 significantly inhibits HBV-driven tumor cells’ growth and migration in vitro and in vivo. HBx-ALKBH5 may form a positive-feedback loop to involve in the HBV-induced liver carcinogenesis, and targeting the loop at ALKBH5 may provide a potential way for HBV-HCC treatment. The online version contains supplementary material available at 10.1186/s12885-021-08449-5.
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影响因子：
64.5
作者：
Cancer Genome Atlas Research Network. Electronic address: wheeler@bcm.edu;Cancer Genome Atlas Research Network
通讯作者：
Cancer Genome Atlas Research Network
影响因子：
5.8
作者：
Bubie A;Zoulim F;Testoni B;Miles B;Posner M;Villanueva A;Losic B
通讯作者：
Losic B
DOI：
10.1007/978-1-4939-6700-1_2
发表时间：
2017-01-01
期刊：
HEPATITIS B VIRUS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ni, Yi;Urban, Stephan
通讯作者：
Urban, Stephan
HBx 蛋白通过稳定 WD 重复结构域 5 蛋白进行 H3K4me3 修饰，从而促进肝癌发生
影响因子：
13.5
作者：
Gao, Weiwu;Jia, Zhengcai;Ni, Bing
通讯作者：
Ni, Bing
影响因子：
11.1
作者：
Nachtergaele S;He C
通讯作者：
He C