Previous studies have demonstrated that perfluorinated chemicals (PFCs) can affect reproduction by disruption of steroidogenesis in experimental animals. However, the underlying mechanism(s) of this disruption remain unknown. Here we investigated the effects and mechanisms of action of 1H, 1H, 2H, 2H-perfluoro-decan-1-ol (
8:2 FTOH) on steroidogenesis using a human
adrenocortical carcinoma cell line (H295R) as a model. H295R cells were exposed to 0, 7.4, 22.2 or 66.6 microM
8:2 FTOH for 24h and productions of
progesterone, 17alpha-OH-progesterone,
androstenedione,
testosterone,
deoxycorticosterone,
corticosterone and
cortisol were quantified by HPLC-MS/MS. With the exception of
progesterone,
8:2 FTOH treatment significantly decreased production of all
hormones in the high dose group. Exposure to
8:2 FTOH significantly down-regulated cAMP-dependent
mRNA expression and
protein abundance of several key steroidogenic
enzymes, including StAR, CYP11A,
CYP11B1,
CYP11B2,
CYP17 and CYP21. Furthermore, a dose-dependent decrease of cellular cAMP levels was observed in H295R cells exposed to
8:2 FTOH. The observed responses are consistent with reduced cellular cAMP levels. Exposure to
8:2 FTOH resulted in significantly less basal (+
GTP) and
isoproterenol-stimulated
adenylate cyclase activities, but affected neither total cellular
ATP level nor basal (-
GTP) or NaF-stimulated
adenylate cyclase activities, suggesting that inhibition of steroidogenesis may be due to an alteration in membrane properties. Metabolites of
8:2 FTOH were not detected by HPLC-MS/MS, suggesting that
8:2 FTOH was not metabolized by H295R cells. Overall, the results show that
8:2 FTOH may inhibit steroidogenesis by disrupting the cAMP signalling cascade.