Sequencing of p53 mutation in established human hepatocellular carcinoma cell line of HHC4 and HHC15 in nude mice.

AIM:To set up cell lines of human hepatocellular carcinoma in nude mice for the research of cell biology and gene therapy.METHODS:Xenotransplantation of human hepatoma into nude mice was carried out and the growth rate, histopathology and immunology of the nude mice were studied. The DNA from xenografts were analyzed by HBV gen and PCR amplification of a fragment of p53 gene exon 7, which were identified by dot blot hybridization, restriction fragments length polymorphism and DNA sequencing.RESULTS:HHC4 and HHC15 cell lines could be successively transplanted in nude mice and the population doubling time was 7 and 5 days respectively. These strains retained the original characteristics of histopathology, secreting AFP and heteroploid karyotypes in human hepatocellular carcinoma. The fragment of HBV gene was detected in the genomic DNA of both hHCC4 and hHCC15, however only HHC4 secreted HBsAg.The mutation at 250 code (C A) and 249 code (G T) were detected respectively in the genomic DNA of HHC4 and HHC15.CONCLUSION:The two cell lines are useful material for the studying of cell biology and gene therapy in human hepatocellular carcinoma and provide molecular biological trace of relationship between high mortality of hepatoma and AFB1 severe pollution of the daily common foods in this district.