Comparative in vitro and in silico study on the estrogenic effects of 2,2-bis(4-chlorophenyl)ethanol, 4,4′-dichlorobenzophenone and DDT analogs

Abstract

DDT and its transformation products (DDTs) are frequently detected in environmental and biological media. Research suggests that DDT and its primary metabolites (DDD and DDE) could induce estrogenic effects by disturbing estrogen receptor (ER) pathways. However, the estrogenic effects of DDT high-order transformation products, and the exact mechanisms underlying the differences of responses in DDT and its metabolites (or transformation products) still remain unknown. Here, besides DDT, DDD and DDE, we selected two DDT high-order transformation products, 2,2-bis(4-chlorophenyl) ethanol (p,p′-DDOH) and 4,4′-dichlorobenzophenone (p,p′-DCBP). We aim to explore and reveal the relation between DDTs activity and their estrogenic effects by receptor binding, transcriptional activity, and ER-mediated pathways. Fluorescence assays showed that the tested 8 DDTs bound to the two isoforms (ERα and ERβ) of ER directly. Among them, p,p′-DDOH exhibited the highest binding affinity, with IC50 values of 0.43 μM and 0.97 μM to ERα and ERβ, respectively. Eight DDTs showed different agonistic activity toward ER pathways, with p,p′-DDOH exhibiting the strongest potency. In silico studies revealed that the eight DDTs bound to either ERα or ERβ in a similar manner to 17β-estradiol, in which specific polar and non-polar interactions and water-mediated hydrogen bonds were involved. Furthermore, we found that 8 DDTs (0.0008–5 μM) showed distinct pro-proliferative effects on MCF-7 cells in an ER-dependent manner. Overall, our results revealed not only for the first time the estrogenic effects of two DDT high-order transformation products by acting on ER-mediated pathways, but also the molecular basis for differential activity of 8 DDTs.