Nitrite-induced deamination and hypochlorite-induced oxidation of DNA in intact human respiratory tract epithelial cells.
No modification of purine or pyrimidine bases was observed when isolated DNA was incubated with 1 mM nitrite at pH 7.4. However, exposure of human bronchial epithelial cells in culture medium at pH 7.4 to nitrite at concentrations of 100 microM or greater led to deamination of purine bases in cellular DNA. Deamination was more extensive in cells exposed to lower extracellular pH values and higher nitrite concentrations. Significant increases in the levels of xanthine and hypoxanthine, putative deamination products of guanine and adenine, respectively, were observed in DNA from nitrite-treated cells but no rise in any base oxidation products such as 8-hydroxyguanine. This pattern of damage suggests that exposure of cells to nitrite (even at pH 7.4) leads to intracellular generation of "reactive nitrogen species" capable of deaminating purines in DNA. In addition, significant DNA strand breakage occurred in nitrite-treated cells. The time course of base damage suggested that the repair of deaminated purine lesions in these cells is slow. By contrast, DNA isolated from cells exposed to hypochlorous acid (HOCl) has significant oxidation of pyrimidine bases and chlorination of cytosine but little oxidation of purines. Exposure of cells to both species (NO(2)(-) plus HOCl) potentiated the oxidative DNA base damage observed but decreased the extent of deamination. We hypothesize that this is due to the formation of nitryl chloride (NO(2)Cl) from reaction of HOCl with *NO(2)(-). The relevance of our observations to events in the stomach and respiratory tract, at sites of inflammation, and in ischemic tissues is discussed.
Spencer
J P
JP
International Antioxidant Research Centre, King's College, London, United Kingdom.
Whiteman
M
M
Jenner
A
A
Halliwell
B
B
D001980:Bronchi / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002461:Cell Line, Transformed; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D003641:Deamination; D004357:Drug Synergism; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006878:Hydroxides / Q000633:toxicity; D006997:Hypochlorous Acid / Q000633:toxicity*; D009573:Nitrites / Q000633:toxicity*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000633:toxicity; D011687:Purines / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism
D006878:Hydroxides; D009573:Nitrites; D011687:Purines; D011743:Pyrimidines; D006997:Hypochlorous Acid; D004247:DNA; C031356:hydroxide ion; D010100:Oxygen