Detection and cloning of oncogenes in the human hepatoma cell-line HCC-M

人肝癌细胞系HCC-M癌基因的检测与克隆

• 批准号: 59440041
• 负责人: TSUCHIYA Masaharu
• 金额: $ 14.4万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1985
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-59440041/
• 关键词: gene transfer; transfection; transformation; hepatoma; NIH; 3T3; hepatitis B virus; DNA; クローニング

We have studied by means of DNA-mediated gene transfer the transforming activity of the DNA of the human hepatoma cell line HCC-M. The following results have been obtained.1. HCC-M possesses characteristics of the cell derived from the human hepatoma.2. It has been demonstrated that HCC-M cells have integrated hepatitis B virus (HBV) DNA although they do not produce HBVassociated antigens.3. DNA from HCC-M induced transformed foci on transfection of NIH/3T3 cells. DNAs from primary transformants were capable of inducing secondary transformants and DNAs from the secondary transformants were also capable of inducing tertiary transformants.4. Most of these secondary transformants were demonstrated to contain both human repetitive sequences and HBV DNA, indicating that the transformants had incorporated exogenous human DNA and HBV DNA as well. This result suggests that there is a close relationship between HBV DNA and transforming gene(s) in HCC-M.5. A 3.5 kd DNA segment was cloned from one of the secondary transformants using pUC13 vector. This segment hybridized with both HBV DNA and v-K-ras genome.From these results it is suggested that the integration of HBV DNA is fairly closely associated with the activation of oncogene(s) in a human hepatoma cell HCC-M.

我们用DNA介导的基因转移方法研究了人肝癌细胞系HCC-M的DNA转化活性。取得了如下结果：1. HCC-M具有人肝癌细胞的特征.研究表明，HCC-M细胞虽然不产生HBV相关抗原，但已整合了B型肝炎病毒（HBV）DNA.来自HCC-M的DNA在转染NIH/3 T3细胞时诱导转化灶。来自初级转化体的DNA能够诱导次级转化体，来自次级转化体的DNA也能够诱导三级转化体。这些次级转化体中的大多数被证明含有人重复序列和HBV DNA，表明转化体也掺入了外源性人DNA和HBV DNA。这一结果提示，在肝癌M型中，HBV DNA与转化基因之间存在着密切的关系。用pUC 13载体从其中一个二级转化子中克隆了一个3.5kd的DNA片段。该片段与HBV DNA和v-K-ras基因组杂交，提示HBV DNA在人肝癌细胞HCC-M中的整合与癌基因的激活密切相关。

项目成果

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