Improved transformation of C3H10T1/2CL8 cells by direct- and indirect-acting carcinogens

Oncogenic transformation of C3H10T1/2CL8 cells was improved by treating the cells 5 days after seeding. Benzo[a]-pyrene-induced transformation was increased 3.5-fold by this method, compared with treating the cells 1 day after seeding. N-Methyl-N'-nitro-N-nitrosoguanidine, which does not transform asynchronous cultures of C3H10T1/2CL8 cells when administered 1 day after seeding, produced on average of 1 focus/dish, with 61% of the dishes exhibiting foci, when administered 5 days after seeding. Propane sultone and aflatoxin B1 also produced marked transformation responses when administered 5 days after seeding. However, 4-dimethylaminoazobenzene did not induce transformation when administered either 1 day or 5 days after seeding. With all chemicals examined, clonal cytotoxicity was reduced when they were administered 5 days after seeding. These results indicate the utility of this new procedure for the qualitative analysis of the transforming ability of chemicals.