Oxidative DNA damage induced by activation of polychlorinated biphenyls (PCBs): implications for PCB-induced oxidative stress in breast cancer.
We have previously reported that mono- and dichlorinated biphenyls (PCBs) can be metabolized to dihydroxy compounds and further oxidized to reactive metabolites which form adducts with nitrogen and sulfur nucleophiles including DNA [Amaro et al. (1966) Chem. Res. Toxicol. 9, 623-629; Oakley et al. (1996) Carcinogenesis 17, 109-114]. The former studies also demonstrated that during the metabolism of PCBs superoxide may be produced. We have therefore examined the abilities of PCB metabolites to induce free radical-mediated oxidative DNA damage using a newly developed, highly sensitive, 32P-postlabeling assay for 8-oxode-oxyguanosine (8-oxodG) [Devanaboyina, U., and Gupta, R. (1996) Carcinogenesis 17, 917-924]. The incubation of 3,4-dichloro-2'5'-dihydroxybiphenyl (100 microM) with calf thymus DNA (300 micrograms/microL) in the presence of the breast tissue and milk-associated enzyme, lactoperoxidase (10 mU/mL), and H2O2 (0.5 mM) resulted in a significant increase in free radical-induced DNA damage (253 8-oxodG/10(6) nucleotides) as compared to vehicle-treated DNA (118 8-oxodG/10(6) nucleotides). Substituting CuCl(2) (100 microM) for lactoperoxidase/H2O2, however, resulted in a substantial increase in 8-oxodG content (2669 8-oxodG/10(6) nucleotides). FeCl(3) was ineffective, suggesting that CuCl(2) but not FeCl(3) mediates oxidation of PCB dihydroxy metabolites, resulting in oxidative DNA damage. The addition of catalase (100 U/mL) and sodium azide (0.1 M) reduced the effect of CuCl(2) (849 and 896 8-oxodG/10(6) nucleotides, respectively), while superoxide dismutase (600 U/mL) moderately stimulated and glutathione (100 microM) substantially stimulated 8-oxodG formation (3014 and 4415 8-oxodG/10(6) nucleotides, respectively). The effect of various buffers as well as the effects of PCB structure on Cu(II)-mediated oxidative DNA damage were examined. These results demonstrate that free radicals and oxidative DNA damage are produced during oxidation of lower chlorinated biphenyls. The relevance of the results is discussed in view of the recent report that increased oxidative DNA base damage is detected in the DNA of human breast tumor tissue.
Oakley
G G
GG
Graduate Center for Toxicology and Preventive Medicine and Environmental Health, University of Kentucky, Lexington 40536, USA.
Devanaboyina
U
U
Robertson
L W
LW
Gupta
R C
RC
D000818:Animals; D001711:Biotransformation; D001943:Breast Neoplasms / Q000139:chemically induced* / Q000378:metabolism*; D002273:Carcinogens / Q000378:metabolism* / Q000633:toxicity*; D002417:Cattle; D002712:Chlorides; D003300:Copper / Q000737:chemistry; D004249:DNA Damage; D005290:Ferric Compounds / Q000737:chemistry; D016166:Free Radical Scavengers / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D007784:Lactoperoxidase / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000502:physiology*; D011078:Polychlorinated Biphenyls / Q000378:metabolism* / Q000633:toxicity*; D013329:Structure-Activity Relationship
D002273:Carcinogens; D002712:Chlorides; D005290:Ferric Compounds; D016166:Free Radical Scavengers; D003300:Copper; D006861:Hydrogen Peroxide; D011078:Polychlorinated Biphenyls; D007784:Lactoperoxidase; C029892:cupric chloride; C024555:ferric chloride