Correlation between O6-methylguanine-DNA-methyltransferase activity and resistance of human cells to the cytotoxic and mutagenic effect of N-methyl-N'-nitro-N-nitrosoguanidine.

Cells from Gardner's syndrome (GS) and familial polyposis coli (FP) patients, persons with a hereditary predisposition to colon cancer, were compared to those of normal persons for sensitivity to the cytotoxic and mutagenic action of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), a model compound chosen because methylating agents have been implicated in colon carcinogenesis. FP cell line GM2355 and GS cell lines 2938 and GM3948 exhibited normal sensitivity to the cytotoxic and mutagenic effects of MNNG. In contrast, GS cell line GM3314 and cells from an apparently normal fetus GM0011 showed extreme sensitivity to the killing and mutagenic effect of this alkylating agent. To determine if the resistance of the various cell lines to MNNG correlated with their ability to remove methyl groups from the O6-position of guanine, we measured their O6-methylguanine-DNA methyltransferase (MT) activity. The resistant cell lines exhibited normal levels of MT; the sensitive strains showed virtually non-detectable levels of this activity. We also compared fibroblasts from a xeroderma pigmentosum (XP) patient (XP12BE, complementation group A), an SV40 virus-transformed XP cell line (XP12ROSV) and a normal cell line transformed by this virus (GM637) for their response to the cytotoxic and mutagenic effect of MNNG and for MT activity. XP12BE cells showed normal sensitivity and a normal level of MT; GM637 cells showed an intermediate level of sensitivity and a reduced level of MT activity; XP12ROSV cells were extremely sensitive to the cytotoxic and mutagenic effect of MNNG and showed virtually non-detectable levels of MT activity. The MT did not remove methyl group from O4-methyl-thymine. These results suggest that O6-methylguanine and/or any other adduct repaired by the methyltransferase, is a potentially cytotoxic and mutagenic lesion. They also indicate that the predisposition to colon cancer of FP and GS patients is not necessarily correlated with an increased sensitivity of their fibroblasts to mutations induced by methylating carcinogens.