Enantioselective Estrogenictty Ofo,p'-dichlorodiphenyltrichloroethane In The Mcf-7 Human Breastrncarcinoma Cell Line

摘要

Research increasingly suggests that selectivity between enantiomers may exist in acute and chronic toxicologieal effects of chiral contaminants. In this study, we used the human breast carcinoma MCF-7 cell line to evaluate enantioselectivity of o,p'-dichlorodiphenyltrichloroethane (o.p'-DDT). Baseline separation of o,p'-DDT enantiomers was achieved on the Chiralcel~R OJ chiral column by high-performance liquid chromatography, and the absolute configuration and optical rotation of the resolved enantiomers were further identified. Significant differences in estrogenic potential were observed between the two enantiomers of o,p'-DDT in the MCF-7 cell proliferation assay (i.e., the E-Scrcen assay) and the real-time quantitative polymerase chain reaction (PCR). In the E-Screen assay, the relative proliferative effect ratios of R-( - )-o,p'-DDT and S-( + )-o,p'-DDT were 89.4 and 27.9%, respectively, and the relative proliferative potency ratios were 0.1 and 0.001%, respectively. Compared to the solvent control. R-( - )-o,p'-DDT induced the maximal increase of 2.31-fold at a concentration of 10~(-6) mol/L, while S-( + )-o,p'-DDT at 10 ~5 mol/L induced the maximal increase of 1,65-fold in estrogenic biomarker pS2 mRNA level. The maximal down-regulation of the transcription levels of estrogen receptor α (ERα) and ERβ by R-(-)-o,p'-DDT were 49 and 40% at the concentration of 10~(-6) mol/L, while those by S-( + )-o,p'-DDT were 24 and 26% at the concentration of 10~(-5) mol/L. The cell proliferation, the up-regulation of pS2. and the down-regulation of ERα and ERβ gene expressions induced by the racemate and enantiomers of o,p'-DDT were all reversed by cotreatment with 10~(-6) mol/L ICI 182,780. Therefore, the enantioselective estrogenicity of o,p'-DDT was likely through the ERa and ERβ signaling pathways. Results from this study suggest the need for considering enantioselectivity of chiral contaminants in chronic ecological toxicities.