Induction of DNA adducts by several polychlorinated biphenyls.
It is known that lower-chlorinated biphenyls are metabolically activated to electrophilic quinoid species capable of binding to DNA. Also, certain metabolites are capable of redox cycling, thereby increasing oxidative stress in biological systems. In the present study, we tested mono-, di-, tri-, tetra-, penta-, hexa-, and heptachlorinated biphenyls for their ability to bind with DNA and to induce oxidative DNA damage. We present additional evidence that several PCB congeners form DNA adducts after metabolic activation, which can be detected by the nuclease P1- or butanol-enrichment procedures of the (32)P-postlabeling technique. Butanol and nuclease P1 enrichments showed different adduct recoveries, depending on the level of chlorination of the biphenyls. Application of the nuclease P1 enrichment showed that the incubation of 2-chloro-; 3, 4-dichloro-; 2,4,4'-trichloro-; 3,4,5-trichloro-; and 2,2',5, 5'-tetrachlorobiphenyl with calf thymus DNA and liver microsomes from rats treated with phenobarbital, followed by oxidation with a peroxidase, produced five to eight different DNA adducts. For these lower-chlorinated biphenyls, butanol enrichment generally showed a lower recovery. For some higher substituted congeners (3,3',4,4', 5-pentachloro-, 2,2',3,4,4',5'-hexachloro-, 2,2',4,4',5, 5'-hexachloro-, and 2,2',3,4,4',5,5'-heptachlorobiphenyl), after butanol enrichment a single dominant spot was observed, which was absent in the nuclease P1 procedure. After incubation of calf thymus DNA with either higher- or lower-chlorinated PCB congeners, we were not able to detect significantly increased levels of oxidative DNA damage above background levels, measured as 8-oxo-7, 8-dihydro-2'deoxyguanosine. In view of the carcinogenicity of PCB mixtures in animals and the ability of PCB metabolites to bind covalently to DNA, rats were orally treated with a mixture of PCBs (Aroclor 1242). PCB-DNA adduct levels were analyzed in PCB target organs: liver, thymus, glandular stomach, spleen, testes, seminal vesicles and prostate DNA. In vivo PCB-DNA adducts could not be detected by either the butanol- or by the NP1-enrichment procedure in rat target tissue DNA. Also, no differences in oxidative DNA damage could be observed between PCB-treated rats and controls. These results indicate a lack of DNA reactivity of PCB mixtures in vivo.
Schilderman
P A
PA
Department of Health Risk Analysis and Toxicology, Maastricht University, Maastricht, The Netherlands.
Maas
L M
LM
Pachen
D M
DM
de Kok
T M
TM
Kleinjans
J C
JC
van Schooten
F J
FJ
D000284:Administration, Oral; D000818:Animals; D001835:Body Weight / Q000187:drug effects; D018736:DNA Adducts / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects*; D007553:Isotope Labeling / Q000379:methods; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects; D010761:Phosphorus Radioisotopes; D011078:Polychlorinated Biphenyls / Q000008:administration & dosage / Q000633:toxicity*; D011467:Prostate / Q000187:drug effects; D051381:Rats; D011917:Rats, Inbred Lew; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000737:chemistry; D013154:Spleen / Q000187:drug effects; D013737:Testis / Q000187:drug effects; D013950:Thymus Gland / Q000187:drug effects; D014018:Tissue Distribution; D018675:Toxicity Tests / Q000379:methods
D018736:DNA Adducts; D010761:Phosphorus Radioisotopes; D011078:Polychlorinated Biphenyls; D015719:Single-Strand Specific DNA and RNA Endonucleases