Tolerance to methylnitrosourea-induced DNA damage is associated with 6-thioguanine resistance in CHO cells

Clones (13 and B) of O6-methylguanine-DNA-methyl-transferase-proficient (MT+) CHO cells showing different levels of resistance to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) but similar MT activity, were found to be sensitive to methyl methanesulphonate and resistant to N-methyl-N-nitrosourea (MNU). A 2.8-fold increase in resistance to MNU-induced cytotoxicity was observed in clone 13 and a 16-fold increase in clone B. A slight increase in survival (1.5-fold) after N-ethyl-N-nitrosourea treatment was observed in clone B. These data indicate that the resistant phenotype is specific for agents that preferentially methylate O atoms in DNA. The survival of MNNG- and MNU-resistant clones as well as of the parental CHO cell line was analysed after exposure to purine analogues substituted in different positions, 8-azaguanine (8-AG), 8-azaadenine (8-AA) and 6-thioguanine (6-TG). A 6-fold increase in resistance to 6-TG was found in clone B, although the hypoxanthine guanine phosphoribosyltransferase gene is functional in these cells. The same cytotoxicity was found in all the lines after treatment with 8-AG and 8-AA. These data are in agreement with the previous observation that clone 13 and clone B belong to two different classes of resistance, clone 13 resistance being explained by MT levels. The finding that clone B is cross-resistant to 6-TG is discussed in the light of a mechanism of tolerance to modifications at specific positions of guanine.