ABSTRACT
Objective To establish a quick method to probe the environmental estrogen-like compounds via fluorescent imaging analysis in high content analysis (HCA) technology.Methods HCA assays were performed to quantitatively in-vestigate the effect of collected environmental pollutants , including bisphenol A , nonylphenol , 1,2-phenylenediamine , 4-aminophenol, resorcinol, 3-aminophenol, 1,4-phenylenediamine and 2,5-diaminotoluene on nuclear granules formation ( Foci-formation) of estrogen receptor α( ERα)-EGFR( enhanced green fluorescent protein ) fusion-protein, which was se-lected as a research model in this study .The results were confirmed by the ERαtranscriptional activity by luciferase as-says.Results Compounds 1,2-phenylenediamine, 4-aminophenol, resorcinol, 3-aminophenol, 1,4-phenylenediamine and 2,5-diaminotoluene sulfate could not induce the ERα-EGFR nuclear granule formation.17-β-Estradiol, bisphenol A, or nonylphenol enhanced ERα-EGFR nuclear granule formation in a dose-dependent manner .The EC50 value was (4.17 ± 0.41) nmol/L, (1.48 ±1.79) μmol/L,or (3.70 ±0.78) μmol/L, respectively.The minimum detectable concentration was 1 nmol/L (17-β-estradiol)and 300 nmol/L (bisphenol A, nonylphenol).In luciferase tests, 17-β-estradiol, bisphe-nol A, or nonylphenol increased ERαtranscriptional activity in a dose-dependent manner ,and the EC50 value was (4.46 ±0.56) nmol/L, (2.31 ±0.21) μmol/L, or (6.60 ±0.94) μmol/L, respectively.The minimum detectable concentration was 3 nmol/L (17-β-estradiol), 300 nmol/L (bisphenol A),and 1 μmol/L (nonylphenol).Conclusion As an efficient method for ERαagonist identification , HCA assays based on the cell image phenotypes analysis can be used in quick recog -nition of environmental compounds with ER agonist-like activity.In all experimental compounds , only bisphenol A and non-ylphenol have a clear ER agonist-like activity .
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Objective To study the related influencing factors of MCF 7 cells proliferation test in vitro to provide some criteria for the standard detection of environmental estrogens (EE) by cell proliferation test of MCF 7 cells in vitro . Methods The sensitivity to E 2 stimulation of different sources of MCF 7 cell lines was detected. Cell proliferation test was performed on the screened sensitive cells in different culture conditions. Results In the estrogen free medium, C cell line of MCF 7 was more sensitive to E 2 stimulation than A and B cell lines ( P
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Objective To observe the estrogenic activity of octylphenol(OP) in vitro and to conduct a preliminary study of its mechanism. Methods The estrogenic activity of OP was detected by cell proliferation test of MCF 7 cells in vitro and the mechanism was preliminarily studied by growth curve analysis, cell cycle analysis, tamoxifen(Tam) antagonistic test and apoptosis detection. Results OP was found to have estrogenic activity to stimulate the proliferation of MCF 7 cells. Cell cycle analysis revealed that the cell proliferation indexes of OP and 17? estradiol(E 2) were higher than those of alcohol. The estrogenic activities of OP and E 2 to stimulate the proliferation of MCR 7 could be antagonized by Tam. Both OP and E 2 could inhibit the cell apoptosis of MCF 7 cells. Conclusion OP possesses estrogenic activity to stimulate the proliferation of MCF 7 cells. The mechanism may be due to binding to the estrogen receptor, which may have effect on cell proliferation and apoptosis.
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Objective To understand the environmental estrogens contamination of tap water and source water. Methods Environmental estrogens in the water samples were concentrated by macroporous adsorption resin H103. The effect of environmental estrogens in concentrated samples was assessed by the recombinant yeast system. Results The results showed that some source water and tap water really contained environmental estrogens. The activity of environmental estrogens in concentrated samples of some source water increased as the volume of concentrated sample increased. The concentrate of 100 ml water sample could show the estrogenic effect. Conclusion Some source waters and tap waters in China have been contaminated by environmental estrogens. Environmental estrogens in source water can not be removed completely by drinking water treatment system of city.
ABSTRACT
Alkylphenols(APs) , the second most widely used commercial non-ionic surfactants in the world, are the main degradated productions of alkylphenol ethoxylates (APEs). The structure, biodegradation, transportation and distribution in environment, environmental monitoring and management are reviewed in this paper. Many meaurement methods of APs show that APs possess estrogenoid effects, that mainly disturb endocrinal and reproductive system, especially, estrogen-dependent organs. APs are associated with decreased fertility, reproductive disfunction, inferbility and cancer formation. APs are able to bind estrogen receptors, to imitate or antagonize estrogen effects, to disturb formation and metabolism of endogenous hormones and receptors. APs are also able to change the structure, quantity and function of cells and to disturb the kinetics of cellcycle at cytological and molecular levels. The more work on epidemiological study and mechanism of APs are needed. Management program for controlling environmental pollution caused by APs is being speedily investigated and put into practice progressively. It is necessary to restrict the use of APEs.