A New Luciferase Reporter Gene Assay for the Detection of Androgenic and Antiandrogenic Effects Based on a Human Prostate Specific Antigen Promoter and PC3/AR Human Prostate Cancer Cells

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  • KIZU Ryoichi
    Graduate School of Natural Science and Technology, Kanazawa University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation
  • OTSUKI Naoki
    Faculty of Pharmaceutical Sciences, Kanazawa University
  • KISHIDA Yoshiko
    Graduate School of Natural Science and Technology, Kanazawa University
  • TORIBA Akira
    Faculty of Pharmaceutical Sciences, Kanazawa University
  • MIZOKAMI Atsushi
    Department of Urology, School of Medicine, Kanazawa University
  • BURNSTEIN Kerry L.
    Department of Molecular and Cellular Pharmacology, University of Miami School of Medicine
  • KLINGE Carolyn M.
    Department of Biochemistry and Molecular Biology, University of Louisville School of Medicine
  • HAYAKAWA Kazuichi
    Graduate School of Natural Science and Technology, Kanazawa University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation

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タイトル別名
  • New Luciferase Reporter Gene Assay for the Detection of Androgenic and Antiandrogenic Effects Based on a Human Prostate Specific Antigen Promoter and PC3 AR Human Prostate Cancer Cells

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We developed a new mammalian cell-based luciferase reporter gene assay for androgenic and antiandrogenic activities of chemicals and environmental samples. Environmental samples usually have a complex matrix that may contain the constituents acting as androgen receptor (AR) agonists, AR antagonists or aryl hydrocarbon receptor (AhR) agonists. AhR agonists are known to elicit the antiandrogenic effect through cross-talk between AR and AhR signal transduction pathways. In this study, PC3/AR human prostate carcinoma cells were transiently transfected with a prostate-specific antigen (PSA) promoter-driven luciferase expression plasmid. The cells were treated with a test compound or an environmental sample for 24 h at 37°C and then measured for luciferase activity. The luciferase activity was induced by dihydrotestosterone (DHT) in a concentration-dependent manner in a concentration range from 10 fM to 1 nM. R1881, a synthetic androgen receptor agonist, induced luciferase activity and its inductive effects was additive to that of DHT. The luciferase activity was not induced by cortisol, a glucocorticoid, progesterone, a progestin, and 17β-estradiol, an estrogen in a concentration range of up to 1 μM. DHT-induced luciferase activity was reduced by bicalutamide and cyproterone acetate, AR antagonists, and also by benzo[a]pyrene, an aryl hydrocarbon receptor agonist, through AhR-mediated pathways. All of these findings indicate that the present assay system correctly responds to AR agonists, AR antagonists and AhR agonist and, therefore, it is a powerful tool for the sensitive and selective screening of chemicals and environmental samples for their androgenic and antiandrogenic activities. We developed the first assay system, in which the expression of luciferase was driven by the promoter of a prostate-specific antigen gene, a typical human androgen-regulated gene.

収録刊行物

  • Analytical Sciences

    Analytical Sciences 20 (1), 55-59, 2004

    社団法人 日本分析化学会

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