[Detection of estrogenic effects of nonylphenol and bisphenol A in vitro reporter gene-based assays].
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.


CONCLUSIONS
The assay we established is usful, NP and BPA showed estrogenic activities.
Zhao
Hai-Tao
HT
Department of Toxicology, Second Military Medical University, Shanghai 200433, China.
Zhang
Tian-Bao
TB
Zhu
Yong-Fei
YF
Wan
Xu-Ying
XY
Zhu
Yuping
Y
Ma
Xili
X
D001559:Benzhydryl Compounds; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D052244:Endocrine Disruptors / Q000633:toxicity*; D004958:Estradiol / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000633:toxicity*; D005260:Female; D017930:Genes, Reporter / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D010636:Phenols / Q000633:toxicity*; D011960:Receptors, Estrogen / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000098826:Bisphenol A Compounds
D001559:Benzhydryl Compounds; D052244:Endocrine Disruptors; D004958:Estradiol; D004968:Estrogens, Non-Steroidal; D010636:Phenols; D011960:Receptors, Estrogen; C025256:nonylphenol; C006780:bisphenol A; D000098826:Bisphenol A Compounds