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The role of integration of HBV DNA into liver tissues in childhood on the development of hepatocellular carcinoma

儿童时期HBV DNA整合到肝组织中对肝细胞癌发生发展的作用

基本信息

批准号：
63570434
负责人：
MORISHIMA Tsuneo
金额：
$ 0.96万
依托单位：
Nagoya University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63570434/
关键词：
Hepatitis B virus Hepatocellular carcinoma Chronic hepatitis HBV DNA Integration Childhood Molecular cloning Southern blot

项目摘要

Numerous epidemiological studies indicated a striking correspondence between areas where frequency of hepatocellular carcinoma (HCC) is high and where hepatitis B virus (HBV) is endemic. Recently, besides serological evidence, the integration of HBV DNA in HCC has been demonstrated using molecular hybridization technique. However, little is known about the integration in HBV carriers, particularly in childhood. Here, we focused on the presence of HBV DNA in liver tissues of HCC, chronic hepatitis and asymptomatic carriers in childhood.The samples were obtained by biopsy and used for DNA extraction. Purified DNA was digested with Hind III, subjected to agarose gel electrophoresis and analyzed by the Southern blot hybridization technique. The age of patients are between 5-15.In 4 of 5 patients of chronic hepatitis, HBV DNA was integrated in the livers. Diffuse hybridization from 5.0 to 20 kb in size was observed, indicating multiple populations of integrated HBV DNA. All of 2 HCC cases (11 yrs old, male and female), HBV DNA was integrated in tissues. However, blot hybridization showed a sharp band in one, showing monoclonal growth of tumor cells, while diffuse smear from 7.6 to 20.0 kb was detected in the other samples. In 2 asymptomatic carriers, HBV DNA integration was also shown in one.Using molecular cloning method, multiple integration pattern was confirmed in the latter case. In the cases of chronic active hepatitis, rearrangement of host-DNA including integrated proviral DNA were found in several patients. This rearrangement may be important for the accivation of cellular oncogene, such as c-myc.From these results, the integration of HBV DNA was completed in early childhood in most, if not all, HBV carriers. For the early prediction of HBV-related HCC, long term follow-up using molecular biological technique may be necessary in HBV carriers, particularly in chronic hepatitis patients.

大量流行病学研究表明，肝细胞癌（HCC）发病率高的地区与乙型肝炎病毒（HBV）流行的地区之间存在惊人的对应关系。最近，除了血清学证据外，利用分子杂交技术证实了HBV DNA在HCC中的整合。然而，对乙肝病毒携带者的整合知之甚少，特别是在儿童中。在这里，我们关注的是HBV DNA在HCC、慢性肝炎和儿童无症状携带者肝组织中的存在。样本通过活组织检查获得，并用于DNA提取。纯化后的DNA经Hind III酶切，琼脂糖凝胶电泳，并用Southern blot杂交技术进行分析。患者年龄在5-15岁之间。在5例慢性肝炎患者中，有4例HBV DNA在肝脏中整合。观察到大小从5.0到20 kb的弥漫性杂交，表明整合的HBV DNA有多个群体。2例HCC患者（11岁，男，女）均有HBV DNA整合到组织中。然而，印迹杂交在其中一个样品中发现了一个尖锐的条带，显示肿瘤细胞单克隆生长，而在其他样品中检测到7.6 - 20.0 kb的弥漫性涂片。在2例无症状携带者中，1例也出现HBV DNA整合。利用分子克隆方法，证实了后者的多重整合模式。在慢性活动性肝炎病例中，在一些患者中发现了包括整合前病毒DNA在内的宿主DNA重排。这种重排可能对细胞癌基因（如c-myc）的激活很重要。从这些结果来看，HBV DNA的整合在大多数（如果不是全部的话）HBV携带者的幼儿期就完成了。对于HBV携带者，特别是慢性肝炎患者，为了早期预测HBV相关HCC，可能需要使用分子生物学技术进行长期随访。