肝脏疾病的患者中的进展感染HBV血清病毒双链线性DNA的比例

StephenW

Gut doi:10.1136/gutjnl-2014-308989

    Hepatology

    Original article

Serum viral duplex-linear DNA proportion increases with the progression of liver disease in patients infected with HBV

    Xing-Liang Zhao1, Jian-Rong Yang2, Sheng-Zhang Lin3, Hui Ma1, Fang Guo1,4, Rui-Feng Yang1, Heng-Hui Zhang1, Jin-Chao Han1, Lai Wei1, Xiao-Ben Pan1

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Author Affiliations

    1Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, P.R. China
    2Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China
    3Department of Hepato-Biliary-Pancreatic Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, P.R. China
    4Department of Microbiology and Immunology, Drexel Institute for Biotechnology and Virology Research, Drexel University College of Medicine, Doylestown, Pennsylvania, USA

    Correspondence to Dr Xiao-Ben Pan, Peking University People's Hospital, Peking University Hepatology Institute; 11# Xizhimeng South Street, Xicheng District, Beijing 100044, P.R. China; [email protected]

    Received 10 December 2014
    Revised 18 May 2015
    Accepted 19 May 2015
    Published Online First 4 June 2015

Abstract

Objective HBV has two forms of genomic DNA, relaxed-circular DNA (rcDNA) and duplex-linear DNA (dlDNA). Compared to rcDNA, dlDNA has been demonstrated to integrate more frequently into host cellular chromosomes, which may have oncogenic consequences. However, the dlDNA proportion relative to total HBV DNA and its clinical significance in patients remain to be investigated.

Design Based on the structural difference between rcDNA and dlDNA, we developed a peptide nucleic acid (PNA)-mediated quantitative real-time PCR (qPCR) clamping assay to measure the proportions of dlDNA in total HBV DNA in sera obtained from patients with chronic hepatitis B (CHB), liver cirrhosis (LC) or LC-developed hepatocellular carcinoma (HCC). The factors that influence the proportion of dlDNA were also investigated.

Results The average dlDNA proportion was approximately 7% in the sera of chronic HBV-infected patients and was elevated in CHB patients with abnormal levels of alanine aminotransferase. The sera dlDNA proportions increased to approximately 14% and 20% in the patients with LC and HCC, respectively. Interferon-α treatment slightly increased the dlDNA proportion in the responders; and nucleotide analogue therapy spuriously elevated the proportion. Moreover, treatment of human hepatoma cells supporting HBV replication with inflammatory cytokines significantly altered the dlDNA proportion in vitro.

Conclusions Using a novel PNA-mediated qPCR clamping assay, we first showed that serum dlDNA proportions progressively increased during the development of HBV-related liver diseases. The dlDNA proportion can be regulated by inflammatory cytokines, suggesting an association among inflammation, increased production of HBV dlDNA and development of HCC.

StephenW

肠道DOI：10.1136 / gutjnl-2014-308989

    肝病

    原创文章

肝脏疾病的患者中的进展感染HBV血清病毒双链线性DNA的比例增加

    兴良Zhao1，建荣Yang2，盛张LIN3，惠MA1，方Guo1,4，锐丰杨1，恒晖Zhang1，金朝Han1，赖伟，肖本PAN1

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作者机构

    1Peking大学人民医院，北京大学肝病研究所，丙型肝炎及免疫治疗肝病，北京，中国公关北京市重点实验室
    临床检验教研室，第一附属医院的温州医学院，温州，浙江，中国公关
    的3Department肝胆胰腺外科，附属第一医院，浙江大学医学院，杭州，浙江，PR中国的
    微生物学与免疫学，德雷塞尔生物技术研究所和病毒学研究，医学德雷塞尔大学学院，宾夕法尼亚州Doylestown，美国4Department

    函授博士晓奔潘，北京大学人民医院，北京大学肝病研究所; 11＃Xizhimeng南大街，西城区，北京100044，中国公关; [email protected]

    收到的2014年12月10日
    修订后的2015年5月18日
    接受2015年5月19日
    网上公布的第一2015年6月4日

抽象

客观HBV有两种形式的基因组DNA，宽松环状DNA（rcDNA）和双工线性DNA（dlDNA）的。相比rcDNA，dlDNA已证明更频繁地整合到宿主细胞的染色体中，可以具有致癌的后果。然而，dlDNA比例相对要总HBV DNA及其临床意义的患者仍有待调查。

设计基于rcDNA和dlDNA之间的结构差异，我们开发肽核酸（PNA）介导的实时定量PCR（定量PCR）检测夹紧衡量dlDNA在从慢性肝炎患者获得总HBV DNA血清中的比例（CHB），肝硬化（LC）或LC-开发肝细胞癌（HCC）。影响dlDNA的比例的因素进行了研究。

结果平均dlDNA比例大约7％的慢性HBV感染患者的血清中和升高慢性乙型肝炎患者的丙氨酸氨基转移酶水平异常。血清dlDNA比例增加至约14％和20％，在LC患者和HCC分别。 α干扰素治疗略有增加，在反应的dlDNA比例;和核苷酸类似物治疗假性升高的比例。此外，治疗肝癌细胞支持HBV的复制与炎性细胞因子的改变显著体外dlDNA比例。

结论：使用一种新型的PNA介导的定量PCR检测夹紧，我们首先发现，血清dlDNA比例乙肝相关肝病的发展过程中逐步增加。该dlDNA比例可由炎症因子调控，提示肝癌的炎症之间的关联，HBV dlDNA增加生产和开发。

君看一叶舟

血清dlDNA比例可以提示病情程度。可以据此开发一种新的监测病情的方法。