Synergism of mutant frequencies in the mouse lymphoma cell mutagenicity assay by binary mixtures of methyl methanesulfonate and ethyl methanesulfonate

Abstract

The effect of mixed mutagen exposures on the rate and type of induced mutants was studied in the L5178Y/TK +/− → TK −/− mouse lymphoma cell mutagenicity assay. In this assay, exposure to ethyl methanesulfonate (EMS) results in more mutants that form large colonies than small colonies. Exposure to methyl methanesulfonate (MMS) results in more mutants that form small colonies than large colonies. Other reports in the literature suggest that large colony TK −/− mutants appear to result from small-scale, perhaps single-gene mutations, and that small-colony TK −/− mutants appear to be associated with chromosomal mutations. Treating cells for 4 h with simple, 2-component mixtures containing 6.45 μg/ml MMS and either 261, 392, 560 or 712 μg/ml EMS resulted in synergism of mutants at each mixture level. The frequencies of total mutants were synergized 12,20,35 and 72%, respectively, in mixed exposures with graded doses of EMS, above the sums of the mixture components. Small colony mutants were synergized to a greater extent than large colony mutants. The frequencies of small colony mutants in mixed exposures were increased 31, 54, 73 and 123%, respectively, while the frequencies of large colony mutants were increased −7, −6, 11 and 39%. Statistical analyses provide strong evidence of synergism (within the limits of the assay) for total and small-colony mutants at all doses of EMS tested, and for large-colony mutants above 400 μg/ml EMS. Similar magnitudes of synergism resulted when other constant levels of MMS (4.30 or 8.60 μg/ml) were mixed with the same graded doses of EMS. The degree of synergism was dependent on EMS concentration but not on MMS concentration.