Activation of ras protooncogenes in 11 2-acetylaminofluorene (AAF)-induced hepatocellular carcinomas (HCCs) or liver cell lines and 9 3'-methyl-(dimethylamino)azobenzene (3'-Me-DAB)-induced HCCs in rats was examined using the NIH3T3 cell transfection assay and oligonucleotide hybridization analysis. Only one cell line established from a AAF-treated rat liver demonstrated transforming activity with a point mutation and ATTA transversion at the second position of H-ras codon 61. The rates of ras activation were thus very low for both AAF- and 3'-Me-DAB-induced rat HCCs, the results thus extending and confirming the findings indicating that ras activation in rat HCCs induced by various type of carcinogens is infrequent.