Evaluation of Time Dependence and Interindividual Differences in Benzo[a]pyrene-Mediated CYP1A1 Induction and Genotoxicity in Porcine Urinary Bladder Cell Cultures∗

∗Sabine Plöttner and Alexandra Borza contributed equally to this work. Jürgen Kuhlmann is deceased.

Abstract

Exposure to tobacco smoke is an established cause of cancer in humans and cigarette smoking is a risk factor for urinary bladder cancer development. Aromatic amines are believed responsible for the bladder-specific carcinogenic effect, but polycyclic aromatic hydrocarbons (PAHs) are also of potential relevance. Urothelial cells contain a number of xenobiotic-metabolizing enzymes, which enable them to convert pro-carcinogens into reactive intermediates. In a preceding study, it was demonstrated using cultured porcine urinary bladder epithelial cells (PUBEC) that CYP1A1 mRNA is induced in a potent manner by treatment with benzo[a]pyrene (BaP). In the present study, the time dependence of these effects was evaluated and whether PUBEC cultures derived from individual donors respond differently to BaP treatment was determined. CYP1A1 induction was analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR), and genotoxic effects were studied using the Comet assay. Incubation of PUBEC with BaP increased CYP1A1 expression and induction of DNA strand breaks in a time-dependent manner. Interindividual differences were found between PUBEC cultures derived from several donor animals with respect to the response to BaP, such that the extent of CYP1A1 induction and magnitude of DNA damage was interrelated. Hence, individual differences in metabolic capacities and responsiveness to xenobiotics of urothelial cells from individual donors may be factors in susceptibility to genotoxic effects induced by PAHs.

This study was supported by a research grant from the Philip Morris External Research Program. We thank Anke Jung, Martina Koch, and Christine Nowak for excellent technical assistance, and Gisela H. Degen and Peter H. Roos for discussion.

Notes

∗Sabine Plöttner and Alexandra Borza contributed equally to this work. Jürgen Kuhlmann is deceased.