High frequency and low specificity of ras gene mutations in rat Zymbal's gland tumors induced by 2-amino-3-methylimidazo[4,5-f]quinoline.

The activating mutations of all three ras genes in rat Zymbal's gland tumors induced by a food mutagen, 2-amino-3-methylimidazo[4,5- f]quinoline (IQ) were analyzed. DNA fragments of the Ha-ras, Ki-ras and N-ras oncogenes were amplified from formalin-fixed and paraffin-embedded tissues by the polymerase chain reaction (PCR) and analyzed for activating mutations involving codons 12, 13 and 61 by oligonucleotide differential hybridization. All nine Zymbal's gland tumors examined, including three papillomas, were found to contain either an Ha-ras or Ki-ras mutation. These mutations were located in either codon 13 or 61 of Ha-ras, and in either codon 12 or 13 of Ki-ras. Of the nine mutations, three were G-->T, three were G-->C, two were G-->A and one was A-->T. Of the nine mutations, eight occurred at guanine bases and seven were transversions. There was no correlation between the types of mutations and the histological types of the tumors. These results suggest that ras gene activation is an important early event in the tumorigenesis induced by IQ in rat Zymbal's gland and that mutations at guanine bases are frequent, though the locations and types of the mutations are not highly specific.