[Detection of estrogenic effects of nonylphenol and bisphenol A in vitro reporter gene-based assays].
Wei Sheng Yan Jiu
; 35(1): 13-5, 2006 Jan.
Article
em Zh
| MEDLINE
| ID: mdl-16598922
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 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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fenóis
/
Genes Reporter
/
Estrogênios não Esteroides
/
Disruptores Endócrinos
Tipo de estudo:
Diagnostic_studies
Limite:
Female
/
Humans
Idioma:
Zh
Revista:
Wei Sheng Yan Jiu
Ano de publicação:
2006
Tipo de documento:
Article