Carcinogenesis, Vol. 21, No. 2, 281-287,
February 2000
© 2000 Oxford University Press
Carcinogenesis |
Transition mutation in codon 248 of the p53 tumor suppressor gene induced by reactive oxygen species and a nitric oxide-releasing compound
Institute of Pharmacology and Toxicology, Rue du Bugnon 27, 1005 Lausanne, Switzerland,
1 Swiss Institute for Experimental Cancer Research, 1066 Epalinges, Switzerland and
2 Chemistry Section, Laboratory of Comparative Carcinogenesis, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, MD 21702, USA
Exposing the human bronchial epithelial cell line BEAS-2B to the nitric oxide (NO) donor sodium 1-(N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA/NO) at an initial concentration of 0.6 mM while generating superoxide ion at the rate of 1 µM/min with the hypoxanthine/xanthine oxidase (HX/XO) system induced C:G
T:A transition mutations in codon 248 of the p53 gene. This pattern of mutagenicity was not seen by `fish-restriction fragment length polymorphism/polymerase chain reaction' (fish-RFLP/PCR) on exposure to DEA/NO alone, however, exposure to HX/XO led to various mutations, suggesting that co-generation of NO and superoxide was responsible for inducing the observed point mutation. DEA/NO potentiated the ability of HX/XO to induce lipid peroxidation as well as DNA single- and double-strand breaks under these conditions, while 0.6 mM DEA/NO in the absence of HX/XO had no significant effect on these parameters. The results show that a point mutation seen at high frequency in certain common human tumors can be induced by simultaneous exposure to reactive oxygen species and a NO source.
Abbreviations: DEA/NO, sodium 1-(N,N-diethylamino)diazen-1-ium-1,2-diolate; DHR 123, dihydrorhodamine 123; ENU, ethylnitrosourea; fish-RFLP/PCR, `fish-restriction fragment length polymorphism/polymerase chain reaction'; HPO, hydroperoxides; HX, hypoxanthine; iNOS, inducible nitric oxide synthase; MDA, malondialdehyde; NO, nitric oxide; NOS, nitric oxide synthases; RH 123, rhodamine 123; ROS, reactive oxygen species; SOD, superoxide dismutase; XO, xanthine oxidase.
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