Deoxyribonucleoside-induced selective modulation of cytotoxicity and mutagenesis.
Treatment of Chinese hamster V79 cells with dThd, dCyd, or dThd plus dCyd increased MNNG-induced AGr-, TGr-, and Ouar-mutant frequencies but did not significantly increase background mutant frequencies. All the AGr colonies that were isolated possessed phenotypes characteristic of HGPRT-deficient mutants, and the deoxyribonucleosides did not selectively affect the growth of the mutants, nor the selecting efficiency of AG, and did not significantly enhance background mutagenesis. These data show that both dThd and dCyd facilitated MNNG-induced mutagenesis. This facilitation was maximal when cells were exposed to the deoxyribonucleosides throughout the first doubling time (24 h) after treatment with MNNG and for 4 more doubling times prior to mutant selection with AG. This indicates that one round of DNA replication was sufficient for mispairing of methylated bases in the DNA with the C and T provided by the deoxyribonucleosides, and that 4-6 doublings prior to mutant selection with AG were necessary to deplete pre-existing hypoxanthine: guanine phosphoribosyl transferase in newly mutated cells. The dCyd facilitated mutagenesis by FdUrd, which was not mutagenic without dCyd, indicating that increased dCTP:dTTP ratios were mutagenic. Treatment with FdUrd plus dCyd also induced FdUrdr cells, suggesting that inhibition of dCyd utilization may prevent the development of FdUrd-resistance in cancer chemotherapy. Although dCyd and dThd facilitated mutagenesis in cells treated with monofunctional alkylating agents that methylate DNA oxygens, facilitation of mutagenesis did not occur in cells treated with BCNU, which cross links DNA, nor with benzo(a)pyrene and aflatoxin B1, which are frame shift mutagens, nor with MMS, which produces barely detectable levels of O-methylation in DNA. Virtually non toxic concentrations of dThd potentiated the cytotoxicity of MNNG more than 10-fold but that of MMS was potentiated only about 2-fold showing that O-alkylation of DNA was associated not only with the facilitation of mutagenesis but also with the potentiation of cytotoxicity. The potentiation of MNNG-induced cytotoxicity was maximal in V79 and L1210 cells after only 2 h treatment with dThd, showing that not even one round of DNA replication was necessary for this potentiation. Moreover, dCyd abolished the potentiation, and, at equitoxic concentrations, MNNG induced higher mutant frequencies than did MMS. These data show that the mechanisms by which methylating agents plus dThd induce mutagenesis are fundamentally different from their mechanisms of cytotoxicity.(ABSTRACT TRUNCATED AT 400 WORDS)
Peterson
A R
AR
Danenberg
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PV
Ibric
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LL
Peterson
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H
D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D001375:Azaguanine / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D004247:DNA / Q000378:metabolism; D003839:Deoxyadenosines / Q000494:pharmacology; D003841:Deoxycytidine / Q000494:pharmacology; D003845:Deoxycytosine Nucleotides / Q000378:metabolism; D003849:Deoxyguanosine / Q000494:pharmacology; D003853:Deoxyribonucleosides / Q000494:pharmacology*; D004351:Drug Resistance; D004357:Drug Synergism; D005467:Floxuridine / Q000494:pharmacology; D007939:Leukemia L1210; D008168:Lung; D008745:Methylation; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D051379:Mice; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation; D013936:Thymidine / Q000494:pharmacology; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors; D013942:Thymine Nucleotides / Q000378:metabolism
D000477:Alkylating Agents; D003839:Deoxyadenosines; D003845:Deoxycytosine Nucleotides; D003853:Deoxyribonucleosides; D009153:Mutagens; D013942:Thymine Nucleotides; D005467:Floxuridine; D003841:Deoxycytidine; D008769:Methylnitronitrosoguanidine; C024107:2'-deoxycytidine 5'-triphosphate; D004247:DNA; D013940:Thymidylate Synthase; D003849:Deoxyguanosine; D001375:Azaguanine; C024157:thymidine 5'-triphosphate; D013936:Thymidine