细胞周期相关激酶介导的病毒宿主的信号，以促进乙型肝炎

StephenW

Gut 2014;63:1793-1804 doi:10.1136/gutjnl-2013-305584

    Hepatology

    Original article

Cell cycle-related kinase mediates viral-host signalling to promote hepatitis B virus-associated hepatocarcinogenesis

    Zhuo Yu 1,2,
    Yue-Qiu Gao 2,
    Hai Feng 3,
    Ying-Ying Lee 4,
    May S Li 3,
    Yuan Tian 4,
    Minnie Y Y Go  4 ,
    Dae-Yeul Yu 5,
    Yue-Sun Cheung 6,
    Paul B S Lai 1,6,
    Jun Yu  1,4,7,
    Vincent W S Wong 1,4,7,
    Joseph J Y Sung  1,7,
    Henry L Y Chan  1,4,7,
    Alfred S L Cheng  3,7

+ Author Affiliations

    1Institute of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China
    2Department of Liver Disease, Shuguang Hospital, Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China
    3School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China
    4Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China
    5Disease Model Research Laboratory, Aging Research Center and World Class Institute, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea
    6Department of Surgery, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China
    7State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, P. R. China

    Correspondence to Professor Alfred Sze-Lok Cheng, School of Biomedical Sciences and State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR 999077, P. R. China; [email protected] and Professor Henry Lik-Yuen Chan, Institute of Digestive Disease, Department of Medicine and Therapeutics and State Key Laboratory of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR 999077, P. R. China; [email protected]

    Received 3 July 2013
    Revised 2 December 2013
    Accepted 23 December 2013
    Published Online First 17 January 2014

Abstract

Background Androgen receptor (AR) signalling contributes to male predominance in hepatocellular carcinoma (HCC), which is more pronounced in HBV-endemic areas. Cell cycle-related kinase (CCRK) is essential for AR-induced hepatocarcinogenesis but its molecular function in HBV-associated HCC remains obscure.

Objective To determine the molecular function of CCRK in HBV-associated HCC.

Design Transcriptional regulation was assessed by chromatin immunoprecipitation, promoter mutation and luciferase reporter assays. Hepatocellular proliferation and tumourigenesis were examined by colony formation, soft agar assays and using HBV X protein (HBx) transgenic mice with low-dose exposure to diethylnitrosamine. Protein expressions were examined in clinical samples and correlated with patient survival by log-rank Mantel–Cox test.

Results Overexpression of CCRK, but not its kinase-defective mutant, activated β-catenin/T cell factor signalling through phosphorylation of glycogen synthase kinase-3β (GSK-3β) at Ser9, led to upregulation of AR transcriptional activity and, subsequently, expression of HBx. The viral transactivator in turn induced CCRK expression through enhanced AR signalling, thus forming a positive regulatory loop. RNA interference silencing of CCRK, which suppressed the CCRK/GSK-3β/β-catenin/AR regulatory loop, significantly suppressed HBx-induced hepatocellular proliferation (p=0.001) and transformation (p<0.001) and remarkably reduced >80% diethylnitrosamine-mediated hepatocarcinogenesis in HBx transgenic mice. Finally, patients with HBV-associated HCC with concordant overexpression of CCRK, GSK-3β phosphorylation at Ser9, active dephosphorylated β-catenin and AR phosphorylation at Ser81 had poorer overall (HR=31.26, p<0.0001) and disease-free (HR=3.60, p<0.01) survival rates.

Conclusions Our findings highlight the critical role of CCRK in a self-reinforcing circuitry that regulates HBV-associated hepatocarcinogenesis. Further characterisation of this intricate viral-host signalling may provide new prognostic biomarkers and therapeutic targets for HCC treatment.

StephenW

肠2014年;63：1793至1804年DOI：10.1136/ gutjnl-2013-305584

    肝病

    原创文章

细胞周期相关激酶介导的病毒宿主的信号，以促进乙型肝炎病毒相关的肝癌

    卓宇1,2，
    跃邱高2，
    海锋3，
    英英肋峨4，
    月子论理3，
    袁体暗4，
    米妮是是围棋4，
    DAE-Yeul玉5，
    岳孙翔6，
    保罗乙对赖1,6，
    君宇1,4,7，
    文森特球法律皇1,4,7，
    约瑟夫Ĵÿ成1,7，
    亨利Ł陈ÿ1,4,7，
    阿尔弗雷德。S L3,7成

+作者所属机构

    消化系统疾病，香港大学中国香港特区，中国大陆的1Institute
    肝病，曙光医院，附属于上海大学中国传统医药，上海，中国的教研室
    生物医学科学3School，中国的香港大学，香港特区，中国大陆
    内科及药物治疗，香港的中国大学，香港特区，中国大陆的4Department
    5Disease模型研究实验室，韩国老龄化研究中心和世界级的研究所，生物科学的韩国研究所和生物技术，大田，共和国
    外科6Department，中国的香港大学，香港特区，中国大陆
    消化系统疾病，香港大学中国香港特区，中国大陆的7State重点实验室

    对应施肇基乐郑教授，生物医学科学和消化系统疾病，香港的中国大学，香港特区999077，中国大陆国家重点实验室学校; [email protected]教授亨利力，陈元，消化系统疾病，内科及药物治疗，消化道疾病，香港的中国大学，香港特区999077，中国大陆国家重点实验室学系研究所; [email protected]

    收到2013年7月3日
    修订后的2013年12月2日
    接受2013年12月23日
    网上公布的第一个2014年1月17日

摘要

背景雄激素受体（AR）信号有助于男性居多的肝细胞癌（HCC），这是乙肝病毒流行地区更为明显。细胞周期相关激酶（CCRK）是必不可少的AR引起的肝癌，但在乙肝相关肝癌的分子的功能尚不清楚。

目的探讨CCRK的乙肝相关肝癌的分子功能。

设计转录调控是由染色质免疫沉淀，子突变和荧光素酶报告基因分析评估。肝细胞增殖和肿瘤发生是由集落形成，软琼脂实验和使用乙肝病毒X蛋白（HBx蛋白）转基因小鼠低剂量接触二乙基亚硝胺检查。蛋白表达进行了检查临床标本和病人的生存用log-rank的Mantel-Cox检验相关。

结果表达CCRK的，而不是它的激酶缺陷型突变体，活化β-连环蛋白/ T细胞因子通过磷酸化糖原合成酶激酶-3β的信令（GSK-3β）在Ser9，导致AR转录活性的上调，且随后，表达的HBx蛋白。该病毒反在通过促进AR信号转致CCRK表达，从而形成了正调节环路。 CCRK，这抑制了CCRK/ GSK-3β/β-连环蛋白/ AR调节环路，显著抑制HBx蛋白诱导的肝细胞增殖（p值=0.001）和变换（对RNA干扰沉默<0.001），显着地减少> 80％diethylnitrosamine-介导的肝癌HBx蛋白在转基因小鼠。最后，例HBV相关HCC与CCRK，GSK-3β的磷酸化在Ser9，主动去磷酸化β-catenin和AR磷酸化Ser81的一致表达了整体较差（HR=31.26，P <0.0001）和无病（HR= 3.60，P <0.01）存活率。

结论：我们的研究结果强调CCRK的一个自我强化的电路，调节乙肝相关肝癌发生中的关键作用。这种复杂病毒的宿主信号的进一步鉴定可以提供新的预后标志物和治疗靶点的肝癌治疗。