Properties of mer- HeLa cells sensitive or resistant to the cytotoxic effects of MNU; effects on DNA synthesis and of post treatment with caffeine.

A line of HeLa cells was shown to be particularly sensitive to N-methyl-N-nitrosurea (MNU) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), but not to a variety of other cytotoxic agents. A resistant line (designated HeLa/A22), was derived by treating HeLa cells repeatedly with MNU. Both the sensitive (HeLa) and resistant (HeLa/A22) cells have a mer- phenotype based both on their reduced rates of loss of O6-methylguanine (O6-MeG) from DNA and their low levels of the enzyme O6-methylguanine methyltransferase (MT). HeLa cells are therefore sensitive to unrepaired O6-MeG in DNA while the HeLa/A22 cells are resistant to unexcised O6-MeG and thus the A22 cells have the mer-rem+ phenotype. MNU produced an immediate dose-dependent inhibition of DNA synthesis in cultures of both sensitive and resistant cells which increased with time until about 4 h after treatment. DNA synthesis then recovered to near control rates in both sensitive and resistant cells before then exhibiting a progressive decrease after about 24 h. DNA synthesis was more depressed at these late times after treatment in cultures of sensitive cells than in those of similarly-treated resistant cells. DNA synthesis remained depressed in sensitive cells but recovered 3 days after treatment in resistant cells. Post treatment incubation of MNU-treated HeLa cells with caffeine did not increase the toxic action of MNU. In contrast, post treatment of the resistant HeLa/A22 cells with caffeine resulted in a dramatic increase in the toxic effects of a higher equitoxic dose of MNU. The depressed rate of DNA synthesis observed in both cell lines after high doses of MNU was partially reversed by post treatment with caffeine in both sensitive and resistant cells. These observations can be interpreted in terms of the effects of caffeine on DNA replication in treated cells.