The role of glutathione in DNA damage by potassium bromate in vitro.
We have investigated the role of reduced glutathione (GSH) in the genetic toxicity of the rodent renal carcinogen potassium bromate (KBrO(3)). A statistically significant increase in the concentration of 8-oxodeoxyguanosine (8-oxodG) relative to deoxyguanosine was measured following incubation of calf thymus DNA with KBrO(3) and GSH or N-acetylcysteine (NACys). This was dependent on these thiols and was associated with the loss of GSH and production of oxidized glutathione. A short-lived (<6 min) intermediate was apparent which did not react with the spin trap dimethylpyrroline N-oxide. DNA oxidation was not evident when potassium chlorate (KClO(3)) or potassium iodate (KIO(3)) were used instead of KBrO(3), though GSH depletion also occurred with KIO(3), but not with KClO(3). Other reductants and thiols in combination with KBrO(3) did not cause a significant increase in DNA oxidation. DNA strand breakage was also induced by KBrO(3) in human white blood cells (5 mM) and rat kidney epithelial cells (NRK-52E, 1.5 mM). This was associated with an apparent small depletion of thiols in NRK-52E cells at 15 min and with an elevation of 8-oxodG at a delayed time of 24 h. Depletion of intra-cellular GSH by diethylmaleate in human lymphocytes decreased the amount of strand breakage induced by KBrO(3). Extracellular GSH, however, protected against DNA strand breakage by KBrO(3), possibly due to the inability of the reactive product to enter the cell. In contrast, membrane-permeant NACys enhanced KBrO(3)-induced DNA strand breakage in these cells. DNA damage by KBrO(3) is therefore largely dependent on access to intracellular GSH.
Parsons
J L
JL
School of Biosciences, The University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Chipman
J K
JK
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D001959:Bromates; D002273:Carcinogens; D002417:Cattle; D002460:Cell Line; D002704:Chlorates / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D020552:Comet Assay; D004247:DNA / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005609:Free Radicals; D005978:Glutathione / Q000378:metabolism / Q000502:physiology*; D019803:Glutathione Disulfide / Q000378:metabolism; D006801:Humans; D007452:Iodates / Q000494:pharmacology; D007668:Kidney / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism; D017680:Potassium Compounds / Q000494:pharmacology; D051381:Rats; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism; D013997:Time Factors
D001959:Bromates; D002273:Carcinogens; D002704:Chlorates; D005609:Free Radicals; D007452:Iodates; D017680:Potassium Compounds; D013438:Sulfhydryl Compounds; C019536:potassium bromate; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine; D005978:Glutathione; C039693:potassium iodate; D019803:Glutathione Disulfide; D000111:Acetylcysteine; C022842:chloric acid