Environmental polycyclic aromatic hydrocarbons affect androgen receptor activation in vitro

Nine structurally different polycyclic aromatic hydrocarbons (PAHs) were tested for their ability to either agonize or antagonize the human androgen receptor (hAR) in a sensitive reporter gene assay based on CHO cells transiently cotransfected with a hAR vector and an MMTV-LUC vector. Benz[a]anthracene (B[a]A), benzo[a]pyrene (B[a]P), fluoranthene, chrysene and 7,12-dimethylbenz[a]anthracene (DMBA) were acting as antiandrogens in vitro, resulting in IC(50) values of 3.2, 3.9, 4.6, 10.3 and 10.4 microM, respectively. Only at the highest concentration tested (10 microM), a slight inhibitory effect by pyrene, phenanthrene, and anthracene was observed. In contrast, dibenzo[a,h]anthracene (DB[a,h]A) gave rise to an agonistic effect, which was added upon the effect of the androgen receptor agonist R1881 (0.1 nM). The antiandrogenic responses by PAHs (10 microM) were found to be fully reversible, determined in the presence of increasing concentrations of R1881. No cytotoxic effects of the tested compounds were observed as determined either by metabolic reduction using AlamarBlue (up to 20 microM) or determined in cells transfected with a constitutively active hAR (up to 10 microM). The well-known ability of certain PAHs to activate the Ah receptor was assessed in H4IIE liver cancer cells, stably transfected with a luciferase reporter gene system. The positive control 2,3,7,8-tetrachlorodibenzodioxin (TCDD) caused a 13-14-fold induction of luciferase activity reaching maximum activity at 0.1 nM. DB[a,h]A, B[a]P, Chrysene, B[a]A and DMBA gave rise to a 4.5-fold induction of luciferase activity at 0.03, 0.4, 0.89, 3.06, and 9.27 microM, respectively, whereas fluoranthene, pyrene, phenanthrene and anthracene were without effect. In conclusion, no clear correlation between the antiandrogenic effects and the Ah receptor activation in vitro was seen. However, the Ah receptor agonists containing four or five aromatic rings (i.e. B [a] A, B [a] P, chrysene, DMBA) appeared to be the most potent antiandrogens (with the exception of DB [a, h] A), whereas those not able to activate the Ah receptor containing three or four aromatic rings (i.e. pyrene, phenanthrene, anthracene) displayed either very weak or no antiandrogenic effect at concentrations up to 10 microM (with the exception of fluoranthene which blocked the hAR at lower concentrations, but did not activate the Ah receptor).