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

• 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
• ISSN: 0340-5761; 1432-0738
• DOI: 10.1007/s00204-003-0437-z

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.