Induction of gene mutation in and cell transformation of mammalian cells by modified purines: 2-aminopurine and 6-N-hydroxylaminopurine

Abstract

2-Aminopurine, a classical mutagen in prokaryotic systems, is inactive as a carcinogen in two animal species. To determine the basis for this discrepancy in the correlation between carcinogenesis and mutagenesis, the ability of 2-aminopurine to induce somatic mutation and neoplastic transformation concomitantly in the same cellular system was examined. 6-N-hydroxylaminopurine, a related modified purine that is a mutagen and a carcinogen, was also studied. 2-Aminopurine was a mutagen in Syrian hamster embryo cells, but its activity was very weak. The maximum induced mutation frequency with either of two mutational markers was only 7 X 10(-6) mutants per surviving cell. 2-Aminopurine also induced morphological transformation of the cells under the same conditions, but the frequency was only approximately 0.04% per surviving colony. Neoplastic transformation of the cells after 2-aminopurine treatment was not observed in these experiments. These results indicate that 2-aminopurine is, at best, a weak transforming agent. The lack of carcinogenic activity in vivo with 2-aminopurine is consistent with these observations. In contrast to the results with 2-aminopurine, 6-N-hydroxylaminopurine was a very effective mutagen in these cells (up to 10(-3) mutants per survivor) and induced morphological transformation of the cells in a dose-dependent manner. Furthermore, neoplastic transformation was induced by this nucleic acid base analog. The correlation of mutagenic activity with transforming ability of these two modified purines supports a relationship between mutagenesis and carcinogenesis. However, relative to other carcinogens, there is a quantitative difference in the ability of 6-N-hydroxylaminopurine to induce cell transformation and mutation. For example, in benzo[a]pyrene-treated cultures, the ratio of the frequency of induced morphological transformation to that of somatic mutation was approximately 100, whereas for 6-N-hydroxylaminopurine-treated cultures, the ratio of transformation to mutation was only 3-12.5. This indicates that 6-N-hydroxylaminopurine is less potent than benzo[a]pyrene in inducing transformation when compared at equal mutagenic potency. This is consistent with our hypothesis that cell transformation, and possibly cancer, occurs predominantly as the result of a mutation at the chromosome level rather than a gene mutation.