Sequence-specific DNA damage induced by carcinogenic danthron and anthraquinone in the presence of Cu(II), cytochrome P450 reductase and NADPH.
The mechanism of metal-mediated DNA damage by carcinogenic danthron (1,8-dihydroxyanthraquinone) and anthraquinone was investigated by the DNA sequencing technique using 32P-labeled human DNA fragments obtained from the human c-Ha-ras-1 protooncogene and the p53 tumor suppressor gene. Danthron caused DNA damage particularly at guanines in the 5'-GG-3', 5'-GGGG-3', 5'-GGGGG-3' sequences (damaged bases are underlined) in the presence of Cu(II), cytochrome P450 reductase and the NADPH-generating system. The DNA damage was inhibited by catalase and bathocuproine, suggesting the involvement of H2O2 and Cu(I). The formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine increased with increasing concentration of danthron. On the other hand, carcinogenic anthraquinone induced less oxidative DNA damage than danthron. Electron spin resonance study showed that the semiquinone radical could be produced by P450 reductase plus NADPH-mediated reduction of danthron, while little signal was observed with anthraquinone. These results suggest that danthron is much more likely to be reduced by P450 reductase and generate reactive oxygen species through the redox cycle, leading to more extensive Cu(II)-mediated DNA damage than anthraquinone. In the case of anthraquinone, its hydroxylated metabolites with similar reactivity to danthron may participate in DNA damage in vivo. We conclude that oxidative DNA damage by danthron and anthraquinone seems to be relevant for the expression of their carcinogenicity.
Ohkuma
Y
Y
Department of Hygiene, Mie University School of Medicine, Tsu, Mie, Japan.
Hiraku
Y
Y
Kawanishi
S
S
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000880:Anthraquinones / Q000494:pharmacology*; D002273:Carcinogens / Q000494:pharmacology*; D002413:Cations, Divalent; D002614:Chelating Agents / Q000494:pharmacology; D003300:Copper / Q000378:metabolism*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000096:biosynthesis; D016166:Free Radical Scavengers / Q000494:pharmacology; D008670:Metals; D009249:NADP / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism*; D010618:Phenanthrolines / Q000494:pharmacology
D000880:Anthraquinones; D002273:Carcinogens; D002413:Cations, Divalent; D002614:Chelating Agents; D016166:Free Radical Scavengers; D008670:Metals; D010618:Phenanthrolines; D009249:NADP; D003300:Copper; D000080242:8-Hydroxy-2'-Deoxyguanosine; C002478:bathocuproine; D009251:NADPH-Ferrihemoprotein Reductase; D003849:Deoxyguanosine; C004315:danthron