Development of a high-performance reporter plasmid for detection of chemicals with androgenic activity

A number of chemicals are present in the environment, and some synthetic chemicals may disrupt endocrine function of wild animals and humans. An effective procedure to screen chemicals for endocrine modulating activity has been needed to ensure the safety of chemicals, and the reporter gene assay te...

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Bibliographic Details
Published in:Archives of toxicology 2003-05, Vol.77 (5), p.274-279
Main Authors: TAKEYOSHI, Masahiro, KUGA, Naoko, YAMASAKI, Kanji
Format: Article
Language:English
Subjects:
Alpha-Globulins - genetics
Androgen Antagonists - classification
Androgen Antagonists - toxicity
Androgens
Animals
Biological and medical sciences
Cercopithecus aethiops
Chemicals
Dose-Response Relationship, Drug
Drug Evaluation, Preclinical - methods
Environmental Pollutants - classification
Environmental Pollutants - toxicity
General aspects. Methods
Genes, Reporter
Humans
Indicators and Reagents
Mammary Tumor Virus, Mouse - genetics
Medical sciences
Plasmids - genetics
Rats
Toxicology
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Summary:A number of chemicals are present in the environment, and some synthetic chemicals may disrupt endocrine function of wild animals and humans. An effective procedure to screen chemicals for endocrine modulating activity has been needed to ensure the safety of chemicals, and the reporter gene assay technique may provide a powerful tool for screening endocrine-disrupting chemicals. We have developed a high-performance reporter plasmid that can trigger high androgen-dependent induction with high selectivity by using mouse mammary tumor virus (MMTV) androgen-responsive elements and a partial fragment of the rat alpha(2u)-globulin promoter region. This new type plasmid can induce higher transcriptional activation than a commercial PGV-P-based construct bearing the SV40 promoter fragment, and the basal induction level of this plasmid is much lower than that of the PGV-P-based construct. Moreover, only androgen derivatives could selectively induce a high response in the reporter gene assay with the new reporter plasmid. This new type of reporter plasmid, ARE-AUG- Luc+, should be of value in endocrine research and in screening to identify endocrine-modulating chemicals.
ISSN:0340-5761
1432-0738
DOI:10.1007/s00204-003-0437-z