Benzo[a]pyrene phenols are more potent inducers of CYP1A1, CYP1B1 and COX-2 than benzo[a]pyrene glucuronides in cell lines derived from the human aerodigestive tract

Abstract

Keratinocyte growth media (KGM) was obtained from Clonetics (San Diego, CA). DMEM and FBS were from Life Technologies (Grand Island, NY). Enzyme immunoassay reagents for assays of prostaglandin E 2 (PGE 2 ) were from Cayman (Ann Arbor, MI). Nitrocellulose membranes were from Schleicher & Schuell (Keene, NH). DNA and RNA were prepared using kits from Qiagen (Chatsworth, CA). The 18S rRNA cDNA was from Ambion (Austin, TX). [a-32 P]dCTP was from Dupont NEN (Boston, MA). Random priming kits were from Boehringer-Mannheim Biochemicals (Indianapolis, IN). B[a]P was from Sigma Chemical (St Louis, MO). 3-OH-B[a]P, 7-OH-B[a]P and 9-OH-B[a]P, as well as 3-benzo[a]pyrenyl-b-d-glucopyranosiduronic acid (B[a]P-3-O-Gluc), 7-benzo[a]pyrenyl-b-d-glucopyranosiduronic acid (B[a]P-7-O-Gluc) and 9-benzo[a]pyrenyl-b-d-glucopyranosiduronic acid (B[a]P-9-O-Gluc) (all used as standards), were obtained from the National Cancer Institute Chemical Abbreviations: B[a]P, benzo[a]pyrene; COX, cyclooxygenase; 3-OH-B[a]P, 3-hydroxy-benzo[a]pyrene; 7-OH-B[a]P, 7-hydroxy-benzo[a]pyrene; 9-OH-B[a]P, 9-hydroxy-benzo[a]pyrene; PGE 2 , prostaglandin E 2 ; UGT, UDPglucuronosyltransferases.