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DNA was extracted from NIH 3T3 cells transformed with DNAs from human primary hepatic cancer (PHC) and Hepatoma 7402 cell line. The transformant DNA was analyzed by Southern transfer and hybridization with ~(32)P-labeled probes of various oncogenes.The EcoRI 7.2 and 9.0 kb bands characteristic of human N-ras gene were identified in transformed NIH 3T3 cells derived both from PHC and 7402 DNA. The BamHI 6.6kb band characteristic of human c-Ha-ras I was present only in 7402 transformants, but not in PHC transformants.Using ~(35)S-methionine incorporation, immunoprecipitation with anti-p21 monoclonal antibodies, SDS-PAGE and autoradiography, it was demonstrated that p21 synthesis was remarkably enhanced in 1402 cells as well as in transformed cells derived from both 7402 and PHC DNA.Taking the data together, it. strongly implies that N-ras is one of the transforming genes for human liver cancer.
DNA was extracted from NIH 3T3 cells transformed with DNAs from human primary hepatic cancer (PHC) and Hepatoma 7402 cell line. The transformant DNA was analyzed by Southern transfer and hybridization with ~ (32) P-labeled probes of various oncogenes. The EcoRI 7.2 and 9.0 kb bands characteristic of human N-ras gene were identified in transformed NIH 3T3 cells derived both from PHC and 7402 DNA. The BamHI 6.6kb band characteristic of human c-Ha-ras I was present only in 7402 transformants, but not in PHC transformants. Using ~ (35) S-methionine incorporation, immunoprecipitation with anti-p21 monoclonal antibodies, SDS-PAGE and autoradiography, it was demonstrated that p21 synthesis was remarkably enhanced in 1402 cells as well as in transformed cells derived from both 7402 and PHC DNA.Taking the data together, it. Strongly implies that N-ras is one of the of the transforming genes for human liver cancer.