Inflammatory mediators accelerate metabolism of benzo[ a ]pyrene in rat alveolar type II cells: The role of enhanced cytochrome P450 1B1 expression

• Published in: Toxicology (Amsterdam) 2013-12, Vol.314 (1), p.30-38
• Main Authors: Šmerdová, Lenka, Neča, Jiří, Svobodová, Jana, Topinka, Jan, Schmuczerová, Jana, Kozubík, Alois, Machala, Miroslav, Vondráček, Jan
• Format: Article
• Language: English
• Subjects: Animals; Aryl Hydrocarbon Hydroxylases - biosynthesis; Aryl Hydrocarbon Hydroxylases - genetics; ATP Binding Cassette Transporter, Sub-Family B - biosynthesis; ATP Binding Cassette Transporter, Sub-Family B - genetics; benzo(a)pyrene; Benzo(a)pyrene - metabolism; Biocompatibility; Blotting, Western; carcinogenicity; Cell Line; Culture Media, Conditioned; CYP1B1; cytochrome P-450; Cytochrome P-450 CYP1B1; Cytochromes P450; Cytokines - metabolism; DNA Adducts; Emergency; etiology; Formations; Genotoxicity; Inflammation; Inflammation Mediators - pharmacology; lung neoplasms; Lungs; macrophages; Metabolism; Metabolites; Oxidoreductases Acting on Aldehyde or Oxo Group Donors - biosynthesis; Oxidoreductases Acting on Aldehyde or Oxo Group Donors - genetics; Polycyclic aromatic hydrocarbons; Pulmonary Alveoli - cytology; Pulmonary Alveoli - drug effects; Pulmonary Alveoli - metabolism; Rats; Real-Time Polymerase Chain Reaction; RNA, Small Interfering; Tandem Mass Spectrometry; toxicology; Transfection; tumor necrosis factor-alpha
• ISSN: 0300-483X; 1879-3185
• DOI: 10.1016/j.tox.2013.09.001

Long-term deregulated inflammation represents one of the key factors contributing to lung cancer etiology. Previously, we have observed that tumor necrosis factor-α (TNF-α), a major pro-inflammatory cytokine, enhances genotoxicity of benzo[a]pyrene (B[a]P), a highly carcinogenic polycyclic aromatic hydrocarbon, in rat lung epithelial RLE-6TN cells, a model of alveolar type II cells. Therefore, we analyzed B[a]P metabolism in RLE-6TN cells under inflammatory conditions, simulated using either recombinant TNF-α, or a mixture of inflammatory mediators derived from activated alveolar macrophage cell line. Inflammatory conditions significantly accelerated BaP metabolism, as evidenced by decreased levels of both parent B[a]P and its metabolites. TNF-α altered production of the metabolites associated with dihydrodiol-epoxide and radical cation pathways of B[a]P metabolism, especially B[a]P-dihydrodiols, and B[a]P-diones. We then evaluated the role of cytochrome P450 1B1 (CYP1B1), which is strongly up-regulated in cells treated with B[a]P under inflammatory conditions, in the observed effects. The siRNA-mediated CYP1B1 knock-down increased levels of B[a]P and reduced formation of stable DNA adducts, thus confirming the essential role of CYP1B1 in B[a]P metabolism under inflammatory conditions. TNF-α also reduced expression of aldo-keto reductase 1C14, which may compete with CYP1B1 for B[a]P-7,8-dihydrodiol and divert it from the formation of ultimate B[a]P dihydrodiol epoxide. Together, the present data suggests that the CYP1B1-catalyzed metabolism of polycyclic aromatic hydrocarbons might contribute to their enhanced bioactivation and genotoxic effects under inflammatory conditions.