title: A versatile transfection assay system to evaluate the biological effects of diverse industrial chemicals.
abstract: 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.
authors.affiliation: Department of Research Planning and Coordination, National Institute of Occupational Safety and Health, 6–21–1 Nagao, Tama-ku, Kawasaki 214–8585, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Koizumi
authors.forename: Shotaro
authors.initials: S
authors.lastname: Ohno
authors.forename: Fuminori
authors.initials: F
authors.lastname: Otsuka
mesh_terms: D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation / Q000187:drug effects*; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D010957:Plasmids; D020218:Response Elements / Q000235:genetics*; D018139:Simplexvirus; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics; D015262:Xenobiotics / Q000633:toxicity*
chemical_list: D014764:Viral Proteins; D015262:Xenobiotics; D013937:Thymidine Kinase