A two-hybrid yeast assay to quantify the effects of xenobiotics on retinoid X receptor-mediated gene expression

Abstract

The aim of this study is to investigate the chemical retinoic acid (RA) disruption at the level of retinoid X receptor (RXR) functioning. This assay makes use of recombined human RXR gene and reporter gene yeast, which specifically expresses β-galactosidase when incubated with exogenous 9- cis retinoic acid (9- cis RA). Agonistic and antagonistic actions of chemicals including a series of phenols, phthalates, organochlorine pesticides (OCPs) were tested in the absence and presence of 5 × 10 −6 mol/L 9- cis RA, at which maximal β-galactosidase activity could be induced. The results obtained reveal that some chemicals, e.g., 2- t-butylphenol, 2-isopropylphenol, 2,4-dichlorophenol (2,4-DCP), 3,4-dichlorophenol (3,4-DCP), 4- tert-octylphenol (4- t-OP) and hexachlorobenzene (HCB), are RXR agonists. Especially, bisphenol A (BPA) showed high induction activity to RXR when tested with metabolization. The 20% relative inhibitory concentration (RIC20) values of r-hexachlorocyclohexane (HCH), p, p′-dichlorodiphenyltrichloroethane ( p, p′-DDT) and 2,4-DCP with metabolization were lower than 1 × 10 −6 mol/L. These results suggest that BPA, HCH, p, p′-DDT and 2,4-DCP are chemicals that pose a threat to hRXR functioning. Altogether the results of the present study show that the newly developed, yeast two-hybrid assay can be used as a valuable tool for identification and quantification of compounds active in disturbing retinoid homeostasis at the level of RXR.