Recognition of environmental estrogen-like compounds based on fluorescent imaging analysis of ERα-EGFP nuclear granules formation
10.7644/j.issn.1674-9960.2015.01.003
- VernacularTitle:基于ERα-EGFP核颗粒形成的荧光成像分析快速识别环境中雌激素样物质
- Author:
Fan FENG
;
Lili WANG
- Publication Type:Journal Article
- Keywords:
high content analysis;
estrogen receptor α;
cellular nuclear granules formation;
environmental estrogen;
transcriptional activity
- From:Military Medical Sciences
2015;(1):12-17
- CountryChina
- Language:Chinese
-
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 .
