Depletion of intracellular glutathione reduces mutations by nitric oxide-donating drugs.
The Mutatect system is a mouse tumor line in which mutations at the hypoxanthine phosphoribosyltransferase (Hprt) locus can be readily detected both in vitro and in vivo. We have previously shown that the nitric oxide-generating drugs, glyceryl trinitrate (GTN) and sodium nitroprusside (SNP), can induce mutations that are readily detected in these cells. In the present report, we have tested the effect of glutathione depletion by buthionine sulfoximine (BSO) on cytotoxicity and mutagenicity by these two drugs. Exposure for 24 h to either drug (123 microM GTN; 500 microM SNP) induced mutations with relatively little cytotoxicity. Pretreatment with 50 microM BSO for 24 h, and then removal at the time of GTN or SNP addition, enhanced cytotoxicity to a modest extent. However, mutagenicity induced by both GTN and SNP was largely abolished. BSO did not affect nitrite accumulation in the medium over a 24-h period, indicating no inhibition of bioactivation of GTN or SNP. Maintaining BSO in the medium for 24 h prior and throughout the period of exposure to GTN or SNP produced a similar effect on mutations. N-Acetylcysteine and oxothiazolidine-4-carboxylate, drugs that are used to increase intracellular glutathione, also blocked mutations. We postulate that a product of the reaction between nitric oxide and intracellular glutathione, such as GSNO or some species derived from it, is promutagenic.
Birnboim
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Ottawa Regional Cancer Centre, University of Ottawa, Ottawa, Ontario, Canada. birnboim@uottawa.ca
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D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D005720:Gamma Rays; D005978:Glutathione / Q000378:metabolism*; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D051379:Mice; D016296:Mutagenesis / Q000187:drug effects* / Q000235:genetics / Q000528:radiation effects; D009153:Mutagens / Q000633:toxicity*; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000037:antagonists & inhibitors / Q000633:toxicity*; D009573:Nitrites / Q000378:metabolism; D005996:Nitroglycerin / Q000037:antagonists & inhibitors / Q000633:toxicity; D009599:Nitroprusside / Q000633:toxicity; D011761:Pyrrolidonecarboxylic Acid; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D014407:Tumor Cells, Cultured
D009153:Mutagens; D020030:Nitric Oxide Donors; D009573:Nitrites; D013844:Thiazoles; D053778:Thiazolidines; D009599:Nitroprusside; D009569:Nitric Oxide; D019328:Buthionine Sulfoximine; D007041:Hypoxanthine Phosphoribosyltransferase; D005996:Nitroglycerin; D005978:Glutathione; D011761:Pyrrolidonecarboxylic Acid; D000111:Acetylcysteine; C029536:2-oxothiazolidine-4-carboxylic acid