Oxidative DNA adducts after Cu(2+)-mediated activation of dihydroxy PCBs: role of reactive oxygen species.
Polychlorinated biphenyls (PCBs) are toxic industrial chemicals, complete carcinogens, and efficacious tumor promoters. However, the mechanism(s) of PCB-mediated carcinogenicity remains largely undefined. One likely pathway by which these agents may play a role in carcinogenesis is the generation of oxidative DNA damage by redox cycling of dihydroxylated PCB metabolites. We have now employed a new (32)P-postlabeling system to examine novel oxidative DNA lesions induced by Cu(2+)-mediated activation of PCB metabolites. (32)P postlabeling of DNA incubated with various PCB metabolites resulted in over a dozen novel polar oxidative DNA adducts that were chromatographically similar for all active agents. The most potent metabolites tested were the hydroquinones (hydroxyl groups arranged para to each other), yielding polar oxidative adduct levels ranging from 55 to 142 adducts/10(6) nucleotides. PCB catechols, or ortho-dihydroxy metabolites, were up to 40% less active than their corresponding hydroquinone congeners, whereas monohydroxylated and quinone metabolites did not produce detectable oxidative damage over that of vehicle. With the exception of 2,4,5-Cl-2',5'-dihydroxybiphenyl, this oxidative DNA damage seemed to be inversely related to chlorine content: no chlorine approximately mono->di->trichlorinated metabolites. Importantly, copper, but not iron, was essential for activation of the PCB metabolites to these polar oxidative DNA adducts, because in its absence or in the presence of the Cu(+)-specific scavenger bathocuproine, no adducts were detected. Intervention studies with known reactive oxygen species (ROS) modifiers suggested that H(2)O(2), singlet oxygen, hydroxyl radical, and superoxide may also be involved in this PCB-mediated oxidative DNA damage. These data indicate a mechanistic role for several ROS, in addition to copper, in PCB-induced DNA damage and provide further support for oxidative DNA damage in PCB-mediated carcinogenesis.
Spencer
Wendy A
WA
James Graham Brown Cancer Center and Department of Pharmacology and Toxicology, University of Louisville, KY 40202, USA.
Lehmler
Hans-Joachim
HJ
Robertson
Larry W
LW
Gupta
Ramesh C
RC
D000818:Animals; D015197:Carcinogenicity Tests; D002274:Carcinogens, Environmental / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D003300:Copper; D004247:DNA / Q000032:analysis*; D018736:DNA Adducts / Q000032:analysis / Q000378:metabolism*; D006873:Hydroquinones / Q000737:chemistry / Q000378:metabolism; D006900:Hydroxylation; D066298:In Vitro Techniques; D006899:Mixed Function Oxygenases / Q000378:metabolism; D009369:Neoplasms / Q000139:chemically induced / Q000175:diagnosis; D018384:Oxidative Stress; D010618:Phenanthrolines / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D011078:Polychlorinated Biphenyls / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D017382:Reactive Oxygen Species; D012474:Salmon / Q000235:genetics*
D002274:Carcinogens, Environmental; D018736:DNA Adducts; D006873:Hydroquinones; D010618:Phenanthrolines; D010761:Phosphorus Radioisotopes; D017382:Reactive Oxygen Species; C042514:cuprous ion; D003300:Copper; D004247:DNA; C002478:bathocuproine; D011078:Polychlorinated Biphenyls; D006899:Mixed Function Oxygenases