DNA DAMAGE BY ALKYLATING AGENTS AND THEIR REPAIR IN HUMAN TUMOR CELLS

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D. Yarosh, R. S. Day, III, and others have shown that approximately 20% of human tumor lines and viral transformed lines are hypersensitive to alkylating agents due to an apparent absence of O6-methylguanine DNA methyltransferase (O6MT); this phenotype has been designated mer-. This enzyme removes alkylation damage at the 0-6 position of guanine but not at other sites in DNA. We and others have isolated O6MT cDNA clones. Our clone has been used as a probe to measure mRNA levels and has also been used to synthesize recombinant protein for antibody development. We have found previously that O6MT mRNA was markedly reduced in all the mer- tumor cells lines examined. The level of O6MT mRNA varied by 10-fold in mer+ cell lines and correlated with known levels of the protein in particular cells. In collaboration with D. Yarosh. O6MT mRNA levels and O6MT activity have now been determined in biopsy specimens of patients with lung and brain tumors. The activity varied widely in tumors from different patients, but was similar in tumor and adjacent normal tissue from the same patient. In general, relative O6MT mRNA levels correlated with activity, but in several lung biopsies the activity was quite low while mRNA levels were not. This indicates that O6MT expression may also be controlled at the translational and/or post-translational levels. Our findings may have important implications in cancer therapy where agents such as BCNU are used. Studies have also been initiated to examine the expression of O6MT in human tumor lines from the panel employed in the NCI drug screening program. The response of the O6MT gene will be studied with other stress-responsive genes in ZO1 CM 07184-04 and will be incorporated into this project.