Oxidation and genotoxicity of fecapentaene-12 are potentiated by prostaglandin H synthase.
The potential importance of prostaglandin H synthase (PGHS) in the genotoxicity of fecapentaene-12 (fec-12) has been indicated by the finding that non-steroidal anti-inflammatory agents (NSAIDs) block the induction of oxidative DNA base damage by fec-12 in HeLa cells. To further investigate the role of PGHS in the metabolic 'activation' and genotoxicity of fec-12, we have measured: (i) oxygen uptake by fec-12 with purified preparations of PGHS; and (ii) the induction of DNA single-strand breaks (SSBs) in HeLa cells exposed to fec-12 in the absence or presence of PGHS inhibitors. Oxygen uptake occurred spontaneously with fec-12 alone but was stimulated 3-fold by the presence of PGHS. The potentiation of fec-12 oxygenation by PGHS was independent of arachidonate and inhibited 45% by indomethacin (INDO). Methylphenylsulphide (MPSI), a reducing substrate expected to compete with fec-12 in peroxidase-dependent co-oxidation reactions, also inhibited PGHS-mediated oxygen uptake with fec-12 by 55%. These results, together with the observation that the inhibitory effects of these agents in combination were additive, suggest that both the cyclooxygenase and peroxidase components of PGHS are involved in the oxidation of fec-12. INDO and MPSI also blocked the induction of DNA SSBs by fec-12 in HeLa cells, indicating that both components of PGHS are also involved in potentiating the genotoxicity of fec-12. It is proposed that this occurs in two ways: firstly, by formation of highly reactive fec-12 hydroperoxides which would generate oxygen radicals through Fenton-like reactions, and secondly, by the generation of oxygen radicals through peroxidase-mediated co-oxidation of fec-12.
Plummer
S M
SM
Centre for Mechanisms of Human Toxicity, Leicester, UK.
Hall
M
M
Faux
S P
SP
D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology*; D004249:DNA Damage; D004357:Drug Synergism; D006367:HeLa Cells; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D007700:Kinetics; D009153:Mutagens / Q000633:toxicity*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010544:Peroxidases / Q000037:antagonists & inhibitors; D011090:Polyenes / Q000633:toxicity*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D013440:Sulfides / Q000494:pharmacology
D016861:Cyclooxygenase Inhibitors; D009153:Mutagens; D011090:Polyenes; D013440:Sulfides; C037357:1-(1-glycero)dodeca-1,3,5,7,9-pentaene; C093850:methylphenylsulfide; D010544:Peroxidases; D011451:Prostaglandin-Endoperoxide Synthases; D007213:Indomethacin