Skip to main content
SpringerLink
Log in

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

  • Published:
Analytical Sciences Aims and scope Submit manuscript

Abstract

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, A. 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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. R. J. Kavlock, G. P. Daston, C. DeRosa, P. Fenner-Crisp, L. E. Gray, Jr., S. Kaattari, G. Lucier, M. Luster, M. J. Mac, C. Maczka, R. Miller, J. Moore, R. Rolland, G. Scott, D. M. Sheehan, T. Sinks, and H. A. Tilson, Environ. Health Perspect., 1996, 104 (Suppl. 4), 715.

    PubMed  PubMed Central  Google Scholar 

  2. R. E. Hester and R. M. Harrison (ed.), “Endocrine Disrupting Chemicals”, 1999, Royal Society of Chemistry, Cambridge.

    Google Scholar 

  3. C. R. Tyler, S. Jobling, and J. P. Sumpter, Crit. Rev. Toxicol., 1998, 28, 319.

    Article  CAS  Google Scholar 

  4. L. E. Gray, Jr., J. Ostby, J. Furr, C. J. Wolf, C. Lambright, L. Parks, D. N. Veeramachaneni, V. Wilson, M. Price, A. Hotchkiss, E. Orlando, and L. Guillette, Human Reproduct. Update, 2000, 7, 248.

    Article  Google Scholar 

  5. D. W. Silversides, C. A. Price, and G. M. Cooke, Can. J. Physiol. Pharmacol., 1995, 73, 1582.

    Article  CAS  Google Scholar 

  6. L. J. Guillette, Jr., D. B. Pickford, D. A. Crain, A. A. Rooney, and H. F. Percival, Gen. Comp. Endocrinol., 1996, 101, 32.

    Article  CAS  Google Scholar 

  7. E. Carlsen, A. Giwercman, N. Keiding, and N. E. Skakkebaek, Br. Med. J., 1992, 305, 609.

    Article  CAS  Google Scholar 

  8. S. H. Swan, E. P. Elkin, and L. Fenster, Environ Health Perspect., 1997, 105, 1228.

    Article  CAS  Google Scholar 

  9. J. Lindzey, M. V. Kumar, M. Grossman, C. Young, and D. J. Tindall, Vitam. Horm., 1994, 49, 383.

    Article  CAS  Google Scholar 

  10. C. A. Quigley, A. DeBille, K. B. Marschke, M. K. El-Awady, E. M. Wilson, and F. S. French, Endocr. Rev., 1995, 16, 271.

    CAS  PubMed  Google Scholar 

  11. W. R. Kelce, C. R. Stone, S. C. Laws, L. E. Gray, J. A. Kempaainen, and E. M. Wilson, Nature, 1995, 375, 581.

    Article  CAS  Google Scholar 

  12. A. M. Vinggaard, E. C. Joergensen, and J. C. Larsen, Toxicol. Appl. Pharmacol., 1999, 155, 150.

    Article  CAS  Google Scholar 

  13. B. Térouanne, B. Tahiri, V. Georget, C. Belon, N. Poujol, C. Avances, F. Orio, Jr., P. Balaguer, and C. Sultan, Mol. Cell. Endocrinol., 2000, 160, 39.

    Article  Google Scholar 

  14. V. S. Wilson, K. Bobseine, C. R. Lambright, and L. E. Gray, Jr., Toxicol. Sci., 2002, 66, 69.

    Article  CAS  Google Scholar 

  15. M. Takeyoshi, N. Kuga, and K. Yamasaki, Arch. Toxicol., 2003, 77, 274.

    Article  CAS  Google Scholar 

  16. N. R. Jana, S. Sarkar, M. Ishizuka, J. Yonemoto, C. Tohyama, and H. Sone, Biochem. Biophys. Res. Commun., 1999, 256, 462.

    Article  CAS  Google Scholar 

  17. A. M. Vinggaard, C. Hnida, and J. C. Larsen, Toxicology, 2000, 145, 173.

    Article  CAS  Google Scholar 

  18. R. Kizu, K. Okamura, A. Toriba, H. Kakishima, A. Mizokami, K. L. Burnstein, and K. Hayakawa, Arch. Toxicol., 2002, 77, 335.

    Article  Google Scholar 

  19. A. Mizokami, A. Gotoh, H. Yamada, E. T. Keller, and T. Matsumoto, J. Urol., 2000, 164, 800.

    Article  CAS  Google Scholar 

  20. P. E. de Ruiter, R. Teuwen, J. Trapman, R. Dijkema, and A. O. Brinkmann, Mol. Cell. Endocrinol., 1995, 110, R1.

    Article  Google Scholar 

  21. J. L. Dai, C. A. Maiorino, P. J. Gkonos, and K. L. Burnstein, Steroids, 1996, 61, 531.

    Article  CAS  Google Scholar 

  22. R. Kizu, K. Okamura, A. Toriba, A. Mizokami, K. L. Burnstein, C. M. Klinge, and K. Hayakawa, Toxicol. Sci., 2003, 76, 299.

    Article  CAS  Google Scholar 

  23. G. M. Cooke, C. A. Price, and R. J. Oko, J. Steroid. Biochem. Mol. Biol., 1998, 67, 347.

    Article  CAS  Google Scholar 

  24. L. E. Gray, Jr., W. R. Kelce, E. Monosson, J. S. Ostby, and L. S. Birnbaum, Toxicol. Appl. Pharmacol., 1995, 131, 108.

    Article  CAS  Google Scholar 

  25. B. L. Roman, R. J. Sommer, K. Shinomiya, and R. E. Peterson, Toxicol. Appl. Pharmacol., 1995, 134, 241.

    Article  CAS  Google Scholar 

  26. N. H. Ing and B. W. O’Malley, “The steroid hormone receptor superfamily”, in “Molecular Endocrinology, Basic Concepts and Clinical Correlations”, ed. B. D. Weintraub, 1995, Raven Press, New York, 195–215.

  27. C. Reyes-Moreno, G. Frenette, J. Boulanger, E. Lavergne, M. V. Govindan, and M. Koutsilieris, Prostate, 1995, 26, 260.

    Article  CAS  Google Scholar 

  28. C. A. Beck, N. L. Weigel, M. L. Moyer, S. K. Nordeen, and D. P. Edward, P. Natl. Acad. Sci. USA, 1993, 90, 4441.

    Article  CAS  Google Scholar 

  29. K. F. Arcaro, P. W. O’Keefe, Y. Yang, W. Clayton, and J. F. Gierthy, Toxicology, 1999, 133, 115.

    Article  CAS  Google Scholar 

  30. J. H. Clemons, L. M. Allan, C. H. Marvin, Z. Wu, B. E. McCarry, D. W. Bryant, and T. R. Zacharewski, Environ. Sci. Technol., 1998, 32, 1853.

    Article  CAS  Google Scholar 

  31. M. Merchant, V. Krishnan, and S. Safe, Toxicol. Appl. Pharmacol., 1993, 120, 179.

    Article  CAS  Google Scholar 

  32. G. Sandblom and E. Varenhorse, Biomed. Pharmacother., 2001, 55, 135.

    Article  CAS  Google Scholar 

  33. J. Toppari, J. C. Larsen, P. Christiansen, A. Giwercman, P. Grandjean, L. J. Guillette Jr., B. Jégou, T. K. Jensen, P. Jouannet, N. Keiding, H. Leffers, J. A. McLachlan, O. Meyer, J. Müller, E. R.-De Meyts, T. Scheike, R. Sharpe, J. Sumpter, and N. E. Skakkebæk, Environ. Health Perspect., 1996, 104 (Suppl. 4), 741

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Graduate School of Natural Science and Technology, Kanazawa University, Kanazawa, 920-0934, Japan

    Ryoichi Kizu, Yoshiko Kishida & Kazuichi Hayakawa

  2. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation, Kawaguchi, 332-0012, Japan

    Ryoichi Kizu & Kazuichi Hayakawa

  3. Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa, 920-0934, Japan

    Naoki Otsuki & Akira Toriba

  4. Department of Urology, School of Medicine, Kanazawa University, Kanazawa, 920-8641, Japan

    Atsushi Mizokam

  5. Department of Molecular and Cellular Pharmacology, University of Miami School of Medicine, Miami, Florida, 33101, USA

    Kerry L. Burnstein

  6. Department of Biochemistry and Molecular Biology, University of Louisville School of Medicine, Louisville, Kentucky, 40292, USA

    Carolyn M. Klinge

Authors
  1. Ryoichi Kizu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Naoki Otsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoshiko Kishida
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Akira Toriba
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Atsushi Mizokam
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kerry L. Burnstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Carolyn M. Klinge
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kazuichi Hayakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ryoichi Kizu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kizu, R., Otsuki, N., Kishida, Y. et al. 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. ANAL. SCI. 20, 55–59 (2004). https://doi.org/10.2116/analsci.20.55

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2116/analsci.20.55

Search

Navigation