Detection and cloning of oncogenes in the human hepatoma cell-line 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。取得了以下成果：1.结论：1.肝细胞癌-M细胞具有人肝癌细胞的特性。已有研究表明，肝细胞癌-M细胞虽然不产生乙肝病毒相关抗原，但仍整合了乙肝病毒DNA。肝细胞癌-M细胞DNA对NIH/3T3细胞的转染率。来自一次转化子的DNA能够诱导二次转化，来自二次转化子的DNA也能够诱导三次转化。大多数二次转化子既含有人的重复序列，又含有HBVDNA，表明转化子中同时含有外源的人DNA和HBVDNA。这一结果提示，HBVDNA与肝癌细胞中的转化基因(S)密切相关。利用pUC13载体从其中一个二级转化子中克隆出一条3.5kd的DNA片段。该片段同时与HBVDNA和v-K-ras基因杂交，提示HBVDNA的整合与人肝癌细胞中癌基因(S)的激活密切相关。

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.