A Versatile Transfection Assay System to Evaluate the Biological Effects of Diverse Industrial Chemicals

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  • Koizumi Shinji
    Department of Research Planning and Coordination, National Institute of Occupational Safety and Health
  • Ohno Shotaro
    Laboratory of Molecular Environmental Health, Department of Life and Health Sciences, Faculty of Pharma-Sciences, Teikyo University
  • Otsuka Fuminori
    Laboratory of Molecular Environmental Health, Department of Life and Health Sciences, Faculty of Pharma-Sciences, Teikyo University

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Gene expression processes are now recognized as important targets of the toxic effects exerted by industrial chemicals. The transient transfection assay is a powerful tool to evaluate such effects. Thus, we developed a versatile assay system by constructing a basic reporter plasmid in which the regulatory DNA sequence to be studied can easily be substituted. To verify the performance of this system, reporter plasmids carrying any of the three distinct regulatory sequences, estrogen responsive element (ERE), glucocorticoid responsive element (GRE) and xenobiotic responsive element (XRE) were constructed. After transfection of human cells, these plasmids successfully expressed the relevant reporter genes in response to specific inducers, β-estradiol, dexamethasone and 3-methylcholanthrene, respectively. Several industrial chemicals were assayed using these reporter plasmids, and the ability of p-dimethylaminoazobenzene to elevate GRE- and XRE-mediated transcription was detected. α-Naphthylamine and o-tolidine were also observed to increase the XRE-mediated response. The transfection assay system established here will be useful to evaluate the effects of a wide variety of industrial chemicals.

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