title: Screening of molecular cell targets for carcinogenic heterocyclic aromatic amines by using CALUX® reporter gene assays.
abstract: Heterocyclic aromatic amines (HCAs) are compounds formed when meat or fish are cooked at high temperatures for a long time or over an open fire. To determine which pathways of toxicity are activated by HCAs, nine out of the ten HCAs known to be carcinogenic in rodents (2-amino-9H-pyrido[2,3-b]indole (AαC), 2-aminodipyrido[1,2-a:3',2-d]imidazole (Glu-P-2), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeAαC), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2)) were tested in the estrogen receptor α (ERα), androgen receptor (AR), glucocorticoid receptor (GR), peroxisome proliferator-activated receptor γ2 (PPARγ2), polycyclic aromatic hydrocarbons (PAH), Nrf2, and p53 CALUX® reporter gene assays. Trp-P-1 was the only HCA that led to a positive response in the ERα, PPARγ2, and Nrf2 CALUX® assays. In the PAH CALUX® assay, Trp-P-2, MeAαC, and AαC induced luciferase activity to a greater extent than MeIQ and PhIP. In the p53 CALUX® assay without a coupled metabolic activation, only Trp-P-1 and Trp-P-2 enhanced luciferase expression; when a metabolic activation step was coupled to the p53 CALUX® assay, Trp-P-1, Glu-P-2, MeIQ, MeIQx, and PhIP induced a positive response. No HCA was positive in the AR and GR CALUX® assays. Taken together, the results obtained show that the battery of CALUX® assays performed in the present study can successfully be used to screen for molecular cell targets of carcinogenic compounds such as HCAs.
authors.affiliation: Institute for Food Toxicology and Analytical Chemistry, University of Veterinary Medicine Hannover, Bischofsholer Damm 15, 30173, Hannover, Germany. pablo.steinberg@tiho-hannover.de.
authors.forename: Pablo
authors.initials: P
authors.lastname: Steinberg
authors.affiliation: BioDetection Systems, Science Park 406, 1098XH, Amsterdam, The Netherlands.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Behnisch
authors.affiliation: BioDetection Systems, Science Park 406, 1098XH, Amsterdam, The Netherlands.
authors.forename: Harrie
authors.initials: H
authors.lastname: Besselink
authors.affiliation: BioDetection Systems, Science Park 406, 1098XH, Amsterdam, The Netherlands.
authors.forename: Abraham A
authors.initials: AA
authors.lastname: Brouwer
mesh_terms: D000588:Amines / Q000633:toxicity*; D000818:Animals; D001681:Biological Assay / Q000379:methods; D002273:Carcinogens / Q000633:toxicity*; D017930:Genes, Reporter / Q000235:genetics*; D006571:Heterocyclic Compounds / Q000633:toxicity*; D008460:Meat / Q000032:analysis*; D051379:Mice; D051381:Rats
chemical_list: D000588:Amines; D002273:Carcinogens; D006571:Heterocyclic Compounds
keywords: CALUX® reporter gene assays; Carcinogens; Heterocyclic aromatic amines