Evaluation of benzo(a)pyrene-induced DNA damage in human endothelial cells using alkaline single cell gel electrophoresis.
The alkaline version of the 'comet assay' was used to evaluate DNA damage in human umbilical vein endothelial cells (HUVEC) exposed to 0.1, 1.0, or 10 microM benzo(a)pyrene for 90min. The genotoxicity was monitored in HUVEC pretreated with the Ah-receptor agonist beta-naphthoflavone (BNF), previously shown to induce cytochrome P4501A1 (CYP1A1) activity in these cells, and in vehicle-treated HUVEC with only constitutive levels of this enzyme. Increased DNA damage was observed only in cells that had been exposed to 10 microM benzo(a)pyrene, cells exposed to BNF being subjected to the most extensive damage. The CYP1A/B-inhibitor alpha-naphthoflavone (ANF) reduced the benzo(a)pyrene-induced DNA-damage in the BNF-treated HUVEC to the same level as in the uninduced cells. The fact that benzo(a)pyrene induced DNA damage in vehicle-treated HUVEC suggests that there may be at least one alternative route of bioactivation for benzo(a)pyrene in these cells. Consequently, judging from the present results it seems as if tobacco-related polycyclic aromatic hydrocarbons (PAHs) may disrupt the function of the endothelial lining in blood vessels with low monooxygenase activity. It is proposed that exposure to Ah receptor agonists via, for example, tobacco smoke, may enhance the DNA-damaging effects of smoke-related genotoxic PAHs in human endothelial cells. The role of PAHs in endothelial dysfunction of tobacco smokers should therefore be further studied.
Annas
A
A
Department of Pharmaceutical Biosciences/Toxicology, Uppsala University, Box 594, SE-751 24 Uppsala, Sweden.
Brittebo
E
E
Hellman
B
B
D000818:Animals; D001564:Benzo(a)pyrene / Q000633:toxicity*; D001571:Benzoflavones / Q000494:pharmacology; D002274:Carcinogens, Environmental / Q000633:toxicity*; D002460:Cell Line; D020552:Comet Assay / Q000379:methods*; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis; D004249:DNA Damage / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000187:drug effects*; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D051379:Mice; D008810:Mice, Inbred C57BL; D009153:Mutagens / Q000633:toxicity*; D019324:beta-Naphthoflavone / Q000494:pharmacology
D001571:Benzoflavones; D002274:Carcinogens, Environmental; D009153:Mutagens; D001564:Benzo(a)pyrene; C011512:alpha-naphthoflavone; D019324:beta-Naphthoflavone; D019363:Cytochrome P-450 CYP1A1