Tsuei D J, Hsu T Y, Chen J Y, Chang M H, Hsu H C, Yang C S
Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Taipei, Republic of China.
J Med Virol. 1994 Mar;42(3):287-93. doi: 10.1002/jmv.1890420316.
The DNA of tumor tissue K1 obtained at autopsy from a case of hepatocellular carcinoma (HCC) in a 9-year-old boy contained integrated hepatitis B virus (HBV) DNA at a single site in the chromosome (case 2, Chang et al.: Hepatology 13:316-320, 1991). To characterize further the integrated viral DNA sequences, a genomic library of the K1 DNA was constructed in the lambda L47.1 vector. One phage clone, designated KTM-1, containing integrated HBV DNA and cellular flanking sequences was obtained from this library. The restriction map and DNA sequence of this clone showed that the integrated HBV DNA was partially deleted and rearranged. The most conserved viral DNA sequences were surface and X genes and arranged in the opposite orientation. The viral core gene was not present. Using chloramphenicol acetyltransferase (CAT) assay, the C-terminal truncated X open reading frame was demonstrated to retain its trans-activating ability. The result suggested that the functional integrated X gene may play a role in hepatocarcinogenesis. The study also showed that the right cellular flanking sequences were human alphoid repetitive sequences.
在一名9岁男孩的肝细胞癌（HCC）尸检中获得的肿瘤组织K1的DNA，在染色体的单个位点含有整合的乙型肝炎病毒（HBV）DNA（病例2，Chang等人：《肝脏病学》13:316 - 320，1991）。为了进一步表征整合的病毒DNA序列，构建了以λL47.1载体为基础的K1 DNA基因组文库。从该文库中获得了一个噬菌体克隆，命名为KTM - 1，其包含整合的HBV DNA和细胞侧翼序列。该克隆的限制性图谱和DNA序列表明，整合的HBV DNA部分缺失并发生了重排。最保守的病毒DNA序列是表面基因和X基因，且排列方向相反。病毒核心基因不存在。使用氯霉素乙酰转移酶（CAT）测定法，证明C末端截短的X开放阅读框保留了其反式激活能力。该结果表明功能性整合的X基因可能在肝癌发生中起作用。该研究还表明，右侧细胞侧翼序列是人类α卫星重复序列。
