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
| Autor: | Yoshiko Kishida, Atsushi Mizokami, Akira Toriba, Naoki Otsuki, Kazuichi Hayakawa, Kerry L. Burnstein, Carolyn M. Klinge, Ryoichi Kizu |
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| Rok vydání: | 2004 |
| Předmět: |
Agonist
Male Bicalutamide medicine.drug_class urologic and male genital diseases Analytical Chemistry chemistry.chemical_compound Genes Reporter Cell Line Tumor medicine Humans Luciferase Luciferases Promoter Regions Genetic biology Chemistry Cyproterone acetate Androgen Antagonists Prostate-Specific Antigen Aryl hydrocarbon receptor Molecular biology Androgen receptor Dihydrotestosterone biology.protein Androgens Signal transduction hormones hormone substitutes and hormone antagonists medicine.drug |
| Zdroj: | Analytical sciences : the international journal of the Japan Society for Analytical Chemistry. 20(1) |
| ISSN: | 0910-6340 |
| Popis: | 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 degrees 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 17beta-estradiol, an estrogen in a concentration range of up to 1 microM. 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. |
| Databáze: | OpenAIRE |
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