Screening of some anti-progestin endocrine disruptors using a recombinant yeast based in vitro bioassay

被引：31
作者：
Chatterjee, Shamba [1 ]
Kumar, Vikas [1 ]
Majumder, Chandrajeet B. [2 ]
Roy, Partha [1 ]
机构：
[1] Indian Inst Technol, Dept Biotechnol, Mol Endocrinol Lab, Roorkee 247667, Uttar Pradesh, India
[2] Indian Inst Technol, Dept Chem Engn, Fluid Particle Res Lab, Roorkee 247667, Uttar Pradesh, India
来源：
TOXICOLOGY IN VITRO | 2008年 / 22卷 / 03期
关键词：
Saccharomyces cerevisiae; progesterone receptor; progestative compounds; fluorescence; endocrine disruptors;
D O I：
10.1016/j.tiv.2007.12.006
中图分类号：
R99 [毒物学（毒理学）];
学科分类号：
100405 ;
摘要：
The present study was aimed to develop a sensitive, fast and user friendly progesterone receptor transactivation assay using recombinant yeast cells, Saccharomyces cerevisiae, modified to express human progesterone receptor (PR) and progesterone response element (PRE) driving the expression of green fluorescent protein. Stimulation of cells with increasing concentrations of progesterone resulted in significant elevation in fluorescence activity, with the minimum effective dose of progesterone being 0.1 nM.RU486, significantly inhibited progesterone induced transactivation and non-progesterogenic steroids failed to transactivate PR till 10 PM concentrations. About 7 different chemicals (mostly pesticides or their metabolites) like DDT and its metabolites, nonylphenol, endosulfan were screened in this assay system for their role in transactivation and they were all found to be anti-progestative and IC50 values within the range of 3-20 mu M. Further, the assay was used to analyze the endocrine disrupting activity of extracted water samples from leather industries known for their high content of various chemicals and it was found to be rich in anti-progestative compounds. It resulted in about 30% reduction in transactivation. In conclusion, we demonstrated that this yeast based bioassay provides a rapid and robust assay for high throughput screening of (anti)progestative compounds from various sources. (c) 2008 Elsevier Ltd. All rights reserved.
引用
收藏
页码：788 / 798
页数：11
相关论文
共 65 条
[1]   Effects of currently used pesticides in assays for estrogenicity, androgenicity, and aromatase activity in vitro [J].
Andersen, HR ;
Vinggaard, AM ;
Rasmussen, TH ;
Gjermandsen, IM ;
Bonefeld-Jorgensen, EC .
TOXICOLOGY AND APPLIED PHARMACOLOGY, 2002, 179 (01) :1-12
[2]  
[Anonymous], 1989, Molecular Cloning
[3]   A yeast estrogen screen for examining the relative exposure of cells to natural and xenoestrogens [J].
Arnold, SF ;
Robinson, MK ;
Notides, AC ;
Guillette, LJ ;
McLachlan, JA .
ENVIRONMENTAL HEALTH PERSPECTIVES, 1996, 104 (05) :544-548
[4]  
AUSUBEL FM, 1995, CURR PROTOCOLS MOL B
[5]  
Barnhart J, 1997, J SOIL CONTAM, V6, P561, DOI 10.1080/15320389709383589
[6]   Congenital malformations and maternal occupation: A registry based case-control study [J].
Bianchi, F ;
Cianciulli, D ;
Pierini, A ;
Costantini, AS .
OCCUPATIONAL AND ENVIRONMENTAL MEDICINE, 1997, 54 (04) :223-228
[7]   Validation and application of a robust yeast estrogen bioassay for the screening of estrogenic activity in animal feed [J].
Bovee, TFH ;
Bor, G ;
Heskamp, HH ;
Hoogenboom, RLAP ;
Nielen, MWF .
FOOD ADDITIVES AND CONTAMINANTS, 2006, 23 (06) :556-568
[8]   Development of a rapid yeast estrogen bioassay, based on the expression of green fluorescent protein [J].
Bovee, TFH ;
Helsdingen, RJR ;
Koks, PD ;
Kuiper, HA ;
Hoogenboom, RLAP ;
Keijer, J .
GENE, 2004, 325 :187-200
[9]   A new highly specific and robust yeast androgen bioassay for the detection of agonists and antagonists [J].
Bovee, Toine F. H. ;
Helsdingen, Richard J. R. ;
Hamers, Astrid R. M. ;
van Duursen, Majorie B. M. ;
Nielen, Michel W. F. ;
Hoogenboom, Ron L. A. P. .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2007, 389 (05) :1549-1558
[10]   Assessment of environmental endocrine disruptors in bald eagles of the Great Lakes [J].
Bowerman, WW ;
Best, DA ;
Grubb, TG ;
Sikarskie, JG ;
Giesy, JP .
CHEMOSPHERE, 2000, 41 (10) :1569-1574
1234567