Generation of DNA base modification following treatment of cultured murine keratinocytes with benzoyl peroxide.
Benzoyl peroxide (BzPO) is a free radical generating compound that acts as a tumor promoter and progressor in mouse skin. BzPO is cleaved in the presence of copper to produce benzoyloxyl and phenyl radicals. Treatment of mutation reporter plasmids with BzPO and copper yields predominantly single-strand breaks and G-->T transversion mutations. To explore the role of base modifications in the possible mammalian mutagenicity of BzPO the formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG) within the DNA of cultured murine keratinocytes was investigated. Treatment with 10 microM BzPO produced a maximum 3-fold increase in levels of 8-OHdG versus vehicle controls within 1-2 h, with significant levels of 8-OHdG persisting 6 h after initial exposure to BzPO. Pretreatment with the copper chelator bathocuproine disulfonic acid reduced the levels of 8-OHdG generated by BzPO to near background. However, treatment with the iron chelator desferal did not. The stable metabolic product of BzPO benzoic acid was ineffective in producing 8-OHdG. Depletion of cellular glutathione with L-buthionine-(S,R)-sulfoximine increased the amount of BzPO-generated 8-OHdG, while supplementation with glutathione monoethyl ester reduced the number of 8-OHdG molecules formed. Collectively, these results suggest that BzPO at non-cytotoxic concentrations undergoes copper-dependent activation to a reactive product to generate 8-OHdG within cultured murine keratinocytes.
King
J K
JK
Department of Environmental Health Sciences, Johns Hopkins School of Hygiene and Public Health, Baltimore, MD 21205, USA.
Egner
P A
PA
Kensler
T W
TW
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D001585:Benzoyl Peroxide / Q000633:toxicity*; D019328:Buthionine Sulfoximine; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000187:drug effects*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects; D007641:Keratolytic Agents / Q000633:toxicity*; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D010618:Phenanthrolines / Q000494:pharmacology
D004791:Enzyme Inhibitors; D007641:Keratolytic Agents; D010618:Phenanthrolines; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; C028559:bathocuproine sulfonate; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine; D001585:Benzoyl Peroxide