HIGH-RATE OF MULTILOCUS DELETION IN A HUMAN TUMOR-CELL LINE

The nature of recessive mutations at the autosomal locus encoding the purine salvage enzyme adenine phosphoribosyl transferase (APRT) was analyzed in a highly malignant human tumor cell line (the colorectal carcinoma line SW620). Mutant strains resistant to the purine analog 8-azaadenine were obtained in two steps. The first step selection for partial drug resistance produced strains hemizygous for APRT as a result of high frequency loss of one allele. In the second step selection, low frequency base substitutions, small deletions, or insertions produced complete azaadenine resistance. Luria-Delbruck fluctuation analysis of each step of this process indicated that the rate of mutation resulting from allele loss was over 100-fold greater than the rate of mutation resulting from base substitution. There was no reproducible difference in the rate of loss of either of the two APRT alleles even though one maps to a rearranged chromosome. Similarly base substitution rates for the two alleles were not significantly different. Polymorphic loci surrounding APRT on chromosome 16 band q24 were lost together with the selected gene in all isolates while polymorphic loci on the short arm of the chromosome were retained. Thus the high frequency loss of APRT in SW620 appears to be the result of multilocus deletions. SW620 derivatives behaving as heterozygotes were also obtained in the first step selections, but these constituted only 5% of isolates.