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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/1736
Title: Screening of some anti-progestin endocrine disruptors using a recombinant yeast based in vitro bioassay
Authors: Chatterjee S.
Kumar V.
Majumder C.B.
Roy, Partha
Published in: Toxicology in Vitro
Abstract: 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 ?M 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 ?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. © 2008 Elsevier Ltd. All rights reserved.
Citation: Toxicology in Vitro (2008), 22(3): 788-798
URI: https://doi.org/10.1016/j.tiv.2007.12.006
http://repository.iitr.ac.in/handle/123456789/1736
Issue Date: 2008
Keywords: Endocrine disruptors
Fluorescence
Progestative compounds
Progesterone receptor
Saccharomyces cerevisiae
ISSN: 8872333
Author Scopus IDs: 17341895000
57212901109
36793883100
35509207200
Author Affiliations: Chatterjee, S., Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India
Kumar, V., Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India
Majumder, C.B., Fluid Particle Research Laboratory, Department of Chemical Engineering, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India
Roy, P., Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India
Funding Details: This work was supported by Council of Scientific and Industrial Research (CSIR) [37(1200)/04/EMRII], and Department of Biotechnology (BT/PR5374/BCE/08/355/2004), Government of India in the form of research funding to PR from both the funding agencies. SC and VK would like to thank CSIR and MHRD, respectively for providing research fellowship in order to carry out this work. We thank Dr. Anand Bachhawat, Institute of Microbial Technology, Chandigarh, India, for yeast host and vectors, Professor J.J. Brosens, Imperial College London, UK, for progesterone receptor and reporter constructs, Professor A.J. Brown, University of Aberdeen, UK, for yEGFP constructs and Dr. Arun Bandyopadhyay, Indian Institute of Chemical Biology, Kolkata, India, for progesterone receptor antibody. Kind help of Professor Ilpo Huhtaniemi, Imperial College London, UK, with all steroidal and non-steroidal test chemicals is greatly acknowledged.
Corresponding Author: Roy, P.; Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, Uttarakhand, India; email: paroyfbs@iitr.ernet.in
Appears in Collections:Journal Publications [BT]

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