title: A novel ERα-mediated reporter gene assay for screening estrogenic/antiestrogenic chemicals based on LLC-MK2 cells.
abstract: Low concentration of endocrine-disrupting chemicals (EDCs) may lead to serious consequences in animals and human, so it is essential to develop an effective assay for EDCs detection. In this study, we developed a novel ERα-mediated reporter gene assay based on the LLC-MK2 cells by co-transfecting pERE-sv40-Luc, hERα-pcDNA3.1, and pRL-tk. Then we determined 17β-estradiol (E2) and some estrogenic/antiestrogenic chemicals to verify the validity of this assay. Data showed that the assay possesses a concentration-dependent responses to E2 and diethylstilbestrol (DES) from 10(-12 )M to 10(-8 )M with EC(50) 3.4 × 10(-10 )M and 5.9 × 10(-10 )M, and ICI 182,780 completely blocks the luciferase activity induced by 10(-9 )M E2. Bisphenol A (BPA), nonylphenol (NP), genistein (GS), and tamoxifen (TAM) also showed corresponding estrogenic or antiestrogenic activity at test concentrations. All evidences proved that the LLC-MK2 reporter gene assay was specific and sensitive to estrogen receptor (ER) agonistic and antagonistic chemicals.
authors.affiliation: Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University , Chengdu, Sichuan , PR China and.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Huang
authors.forename: Lishi
authors.initials: L
authors.lastname: Zhang
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Zhu
authors.forename: Yun
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D020847:Estrogen Receptor Modulators / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000235:genetics*; D004967:Estrogens / Q000494:pharmacology*; D017930:Genes, Reporter; D008253:Macaca mulatta
chemical_list: D020847:Estrogen Receptor Modulators; D047628:Estrogen Receptor alpha; D004967:Estrogens
keywords: ERα-mediated reporter gene assay; LLC-MK2 cells; estrogenic/antiestrogenic activity; luciferase