Title: Ultraviolet-B-induced oxidative DNA base damage in primary normal human epidermal keratinocytes and inhibition by a hydroxyl radical scavenger.
Authors: Pelle, Edward; Huang, Xi; Mammone, Thomas; Marenus, Kenneth; Maes, Daniel; Frenkel, Krystyna
Published In J Invest Dermatol, (2003 Jul)
Abstract: To evaluate the effects of ultraviolet-induced environmental trauma on human skin cells, primary normal human epidermal keratinocytes were exposed to ultraviolet-B radiation (290-320 nm). We found that relatively low doses of ultraviolet-B (62.5-500 mJ per cm2) caused dose-dependent increases in 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG), a biomarker of oxidative DNA damage. Unirradiated normal human epidermal keratinocytes contained 1.49 (+/- 0.11) 8-oxo-dG per 10(6) 2'-deoxyguanosine (dG) residues in cellular DNA, which increased linearly to as high as 6.24 (+/- 0.85) 8-oxo-dG per 10(6) dG after irradiation with 500 mJ per cm2. Further, this oxidative damage was reduced by 60.7% when the cells were pretreated with 1 mM mannitol. As hydrogen peroxide (H2O2) is known to be generated during oxidative stress, its accumulation in ultraviolet-B-irradiated normal human epidermal keratinocytes was also assessed and correlated to 8-oxo-dG formation. An ultraviolet-B-induced increase in H2O2 was observed in normal human epidermal keratinocytes and its production was inhibited by the addition of catalase. Based on the ability of a neutral molecule like H2O2 to permeate membranes, our data indicate that, after ultraviolet-B irradiation, H2O2 migrates from the cytosol to the nucleus where it participates in a Fenton-like reaction that results in the production of hydroxyl radicals (OH*), which may then cause 8-oxo-dG formation in cellular DNA. This conclusion is supported by our data showing that OH* scavengers, such as mannitol, are effective inhibitors of oxidative DNA base damage. Although increased levels of 8-oxo-dG were previously found in immortalized mouse keratinocytes exposed to ultraviolet-B radiation, we now report the induction of 8-oxo-dG in normal human skin keratinocytes at ultraviolet-B doses relevant to human skin exposure.
PubMed ID: 12839579
MeSH Terms: Antineoplastic Agents/pharmacology; Antioxidants/pharmacology; Cells, Cultured; DNA Damage/drug effects*; Deoxyguanosine/analogs & derivatives*; Deoxyguanosine/metabolism; Diuretics, Osmotic/pharmacology; Epidermis/cytology*; Epidermis/metabolism; Epidermis/radiation effects; Free Radical Scavengers/pharmacology*; Humans; Hydrogen Peroxide/metabolism; Hydroxyl Radical/metabolism; Keratinocytes/cytology; Keratinocytes/metabolism*; Keratinocytes/radiation effects*; Mannitol/pharmacology; Oxidation-Reduction; Salicylic Acids/pharmacology; Thiourea/pharmacology; Ultraviolet Rays; beta Carotene/pharmacology