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Tim Hofer, Carine Badouard, Edyta Bajak, Jean-Luc Ravanat, Åse Mattsson, Ian A. Cotgreave

Hydrogen peroxide causes greater oxidation in cellular RNA than in DNA

Keywords: A549 cells, electrospray ionization, Mass spectrometry, oxidative stress, 8-oxodGuo, 8-oxoGuo

Human A549 lung epithelial cells were challenged with ¹⁸O-labeled hydrogen peroxide ([¹⁸O]-H₂O₂), the total RNA and DNA extracted in parallel, and analyzed for ¹⁸O-labeled 8-oxo-7,8-dihydroguanosine ([¹⁸O]-8-oxoGuo) and 8-oxo-7,8-dihydro-2?-deoxyguanosine ([¹⁸O]-8-oxodGuo) respectively, using high-performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-MS/MS). [¹⁸O]-H₂O₂ exposure resulted in dose-response formation of both [¹⁸O]-8-oxoGuo and [¹⁸O]-8-oxodGuo and ¹⁸O-labeling of guanine in RNA was 14–25 times more common than in DNA. Kinetics of formation and subsequent removal of oxidized nucleic acids adducts were also monitored up to 24 h. The A549 showed slow turnover rates of adducts in RNA and DNA giving half-lives of approximately 12.5 h for [¹⁸O]-8-oxoGuo in RNA and 20.7 h for [¹⁸O]-8-oxodGuo in DNA, respectively.

Biological Chemistry, Walter de Gruyter

Print ISSN: 1431-6730
Volume: 386, 04/2005
Pages: 333 - 337

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