title: Effects of cigarette smoke condensate on primary urothelial cells in vitro.
abstract: Cigarette smoking is a risk factor for bladder cancer. Since urothelial cells express phase I and II enzymes these cells are able to metabolize precarcinogens into DNA reactive intermediates. Cigarette smoke is a complex mixture containing at least 80 known carcinogens. In this context especially aromatic amines and polycyclic aromatic hydrocarbons are discussed as being responsible for bladder-carcinogenicity. Cell cultures of primary porcine urinary bladder epithelial cells (PUBEC) have been useful models for studies on bladder-specific effects. These cells are metabolically competent and found to be a valuable tool for examining effects of cigarette smoke constituents. In the present study PUBEC were utilized to investigate the effects of the complex mixture cigarette smoke condensate total particulate matter (CSC TPM) with emphasis on induction of cytochrome P-450 1A1 (CYP1A1) and genotoxic effects. CYP1A1 induction was investigated by Western blot and flow cytometry. The most pronounced effects were found after 24 h of incubation with 1-10 μg/ml CSC TPM. Maximal induction was observed at 5 μg/ml by flow cytometry and at 10 μg/ml by Western blot analysis. Genotoxic effects were investigated by means of alkaline single-cell gel electrophoresis ("comet assay") with and without the use of the DNA repair enzyme formamidopyrimidine-DNA glycosylase (Fpg) and the micronucleus (MN) test. A numerical concentration-dependent increase in Fpg-sensitive sites indicating oxidative DNA damage and a quantitative rise in MN formation were noted. The CSC utilized in this study contained low amounts of benzo[a]pyrene, 4-aminobiphenyl, and 2-naphthylamine. With regard to the observed CYP1A1 induction, these substances cannot explain the CYP1A1 inducing effect of CSC TPM. It is possible that other compounds within CSC TPM contribute to CYP1A1 induction in our cellular model.
authors.affiliation: Leibniz Research Centre for Working Environment and Human Factors (IfADo), Dortmund, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Plöttner
authors.forename: Claudia
authors.initials: C
authors.lastname: Behm
authors.forename: Hermann M
authors.initials: HM
authors.lastname: Bolt
authors.forename: Wolfram
authors.initials: W
authors.lastname: Föllmann
mesh_terms: D000393:Air Pollutants / Q000633:toxicity*; D000818:Animals; D002478:Cells, Cultured; D020552:Comet Assay; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis*; D004249:DNA Damage / Q000187:drug effects*; D043382:DNA-Formamidopyrimidine Glycosylase / Q000378:metabolism; D004790:Enzyme Induction; D005434:Flow Cytometry; D015151:Immunoblotting; D015162:Micronucleus Tests; D013552:Swine; D014028:Tobacco Smoke Pollution / Q000009:adverse effects*; D001743:Urinary Bladder / Q000166:cytology / Q000187:drug effects*; D019459:Urothelium / Q000166:cytology
chemical_list: D000393:Air Pollutants; D014028:Tobacco Smoke Pollution; D019363:Cytochrome P-450 CYP1A1; D043382:DNA-Formamidopyrimidine Glycosylase