乙型肝炎病毒-Alu-聚合酶链反应检测肝细胞癌中乙型肝炎病毒DNA的整合  

Detection of hepatitis B virus DNA integration in hepatocellular carcinoma using hepatitis B virus-Alu-polymerase chain reaction

作　　者：解俊侠[1] 张亚飞[1] 谢琴秀[1] 郜玉锋[1] 胡立芬[1] 李家斌[1] 李旭[1] 

机构地区：[1]安徽医科大学第一附属医院感染科,安徽合肥230032

出　　处：《中华传染病杂志》2008年第4期235-239,共5页Chinese Journal of Infectious Diseases

基　　金：基金项目：国家自然科学基金资助项目（30471518）

摘　　要：目的了解肝细胞癌（HCC）中HBVDNA的整合情况。方法收集24例HBsAg阳性患者的HCC组织，提取其DNA，应用套式PCR原理，在设计的引物中引入U碱基，根据已知基因序列和人Alu重复序列分别设计引物，建立克隆整合的HBVDNA及其相邻的细胞基因序列的PCR技术。PCR产物测序所获结果经美国国家生物技术信息中心（NCBI）BLAST及MapViewer检索确定HBV整合在染色体上的精确位置。结果24份HCC组织中，14份存在HBV整合现象，整合的标本中11份正向插入宿主基因，8份反向插入宿主基因，其中5份标本既有正向插入也有反向插入，从病毒基因分析，整合可发生于X基因的任何长度，且均以截短形式插入宿主细胞DNA。另有8份标本未测到整合。结论HBV在HCC细胞染色体上的整合呈不均衡分布。Objective To detect the integration of hepatitis B virus （HBV） DNA in HBV-related human hepatocellular carcinomas （HCC）. Methods Extracted DNA from the liver tissue samples and amplified by nested polymerase chain reaction （PCR） with specially designed U-base primers. According to the known genes and human Alu repeat sequences （Alu repeat）, primers were designed respectively. Integrated clones combined target HBV DNA and the adjacent cell gene sequences were established by PCR and products were sequenced by biotechnology companies. Accurate locations of HBV genes integrated in the human genomes were analyzed by national center for biotechnology information （NCBI） basic local alignment search tool （BLAST） and Map Viewer search. Results In 24 HBsAg positive HCC samples, 15 cases showed the integrations of HBV fragment. And the other 8 samples didn＇t show any evidence of integration. Among 14 samples with integration, forward insertions of HBV DNA into the host chromosomal DNA were found in 10 samples and reverse insertions were found in 8 samples while both forward and reverse insertions were found in 5 samples. Analysis from viral-cellular junctions suggested that the integrations were all happened with truncated viral DNA and could be in any locus of X gene. Conclusion HBV DNA integration is not distributed evenly throughout the host genome.

关 键 词：Alu元件 肝炎病毒 乙型 病毒整合 癌 肝细胞 DNA 病毒 

分 类 号：R512.62[医药卫生—内科学] R735.7[医药卫生—临床医学]