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

Research Project

Project/Area Number	59440041
Research Category	Grant-in-Aid for General Scientific Research (A)
Allocation Type	Single-year Grants
Research Field	Gastroenterology
Research Institution	Keio University
Principal Investigator	TSUCHIYA Masaharu 慶応義塾大学, 医, 教授 (60051124)
Project Period (FY)	1984 – 1985
Project Status	Completed (Fiscal Year 1985)
Budget Amount *help	¥22,500,000 (Direct Cost: ¥22,500,000) Fiscal Year 1985: ¥3,500,000 (Direct Cost: ¥3,500,000) Fiscal Year 1984: ¥19,000,000 (Direct Cost: ¥19,000,000)
Keywords	gene transfer / transfection / transformation / hepatoma / NIH / 3T3 / hepatitis B virus / DNA / クローニング
Research Abstract	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.