Title: [Detection of estrogenic effects of nonylphenol and bisphenol A in vitro reporter gene-based assays].
Abstract:Objective To explore the estrogenic effects and disruptive mechanism of NP and BPA by reporter gene-based assays we developed. Methods pERE-Luc plamid was generated by inserting estrogen response elem...Objective To explore the estrogenic effects and disruptive mechanism of NP and BPA by reporter gene-based assays we developed. Methods pERE-Luc plamid was generated by inserting estrogen response element (ERE) fragment into MCS of pGL3-promoter vector. MCF7 cells were cotransfected with pERE-Luc and phRL-SV40 using Sofast transfection reagent. The cells then treated with 17beta-estradiol (E2), tamoxifen (Tam), nonylphenol(NP) and bisphenol A (BPA) and expression of the repoter gene in the cell lysates was assayed using Dual-Lucferase reporter assay system. Results The pERE-Luc plasmid was constructed. Luciferase activities of MCF7 cells transfected pERE-Luc showed dose-responed realitionship with E2. 1 x 10(-11) mol/L E2 could induce the expression of reporter gene and 1 x 10(-9) mol/L E2 resulted in the largest luciferase activity. E2 couldn't induce the luciferase activity without pERE-Luc. Tam is a complete antagonist, inhibited the E2-induced luciferase expression. NP induced the luciferase activity at concertrations > 1 x 10(-6) mol/L, BPA induced the luciferase activity at concertrations > 1 x 10(-6) mol/L. The estrogenic activity of NP was more than BPA. Conclusion The assay we established is usful, NP and BPA showed estrogenic activities.Read More
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
Indexed In: ['pubmed']
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Cited By Count: 1
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