Growth Inhibition by Testosterone in an Androgen Receptor Splice Variant-Driven Prostate Cancer Model

Prostate. 2016 Dec;76(16):1536-1545. doi: 10.1002/pros.23238. Epub 2016 Jul 30.

Abstract

Background: Castration resistance creates a significant problem in the treatment of prostate cancer. Constitutively active splice variants of androgen receptor (AR) have emerged as drivers for resistance to androgen deprivation therapy, including the next-generation androgen-AR axis inhibitors abiraterone and enzalutamide. In this study, we describe the characteristics of a novel castration-resistant prostate cancer (CRPC) model, designated JDCaP-hr (hormone refractory).

Methods: JDCaP-hr was established from an androgen-dependent JDCaP xenograft model after surgical castration. The expression of AR and its splice variants in JDCaP-hr was evaluated by immunoblotting and quantitative reverse transcription-polymerase chain reaction. The effects of AR antagonists and testosterone on JDCaP-hr were evaluated in vivo and in vitro. The roles of full-length AR (AR-FL) and AR-V7 in JDCaP-hr cell growth were evaluated using RNA interference.

Results: JDCaP-hr acquired a C-terminally truncated AR protein during progression from the parental JDCaP. The expression of AR-FL and AR-V7 mRNA was upregulated by 10-fold in JDCaP-hr compared with that in JDCaP, indicating that the JDCaP and JDCaP-hr models simulate castration resistance with some clinical features, such as overexpression of AR and its splice variants. The AR antagonist bicalutamide did not affect JDCaP-hr xenograft growth, and importantly, testosterone induced tumor regression. In vitro analysis demonstrated that androgen-independent prostate-specific antigen secretion and cell proliferation of JDCaP-hr were predominantly mediated by AR-V7. JDCaP-hr cell growth displayed a bell-shaped dependence on testosterone, and it was suppressed by physiological concentrations of testosterone. Testosterone induced rapid downregulation of both AR-FL and AR-V7 expression at physiological concentrations and suppressed expression of the AR target gene KLK3.

Conclusions: Our findings support the clinical value of testosterone therapy, including bipolar androgen therapy, in the treatment of AR-overexpressed CRPC driven by AR splice variants that are not clinically actionable at present. Prostate 76:1536-1545, 2016. © 2016 Wiley Periodicals, Inc.

Keywords: AR-V7; androgen deprivation therapy; castration-resistant prostate cancer; testosterone therapy.

MeSH terms

  • Androgen Receptor Antagonists / pharmacology
  • Animals
  • Cell Proliferation / drug effects
  • Disease Models, Animal
  • Down-Regulation / drug effects
  • Genetic Variation
  • Heterografts
  • Humans
  • Male
  • Mice
  • Mice, Nude
  • Neoplasm Transplantation
  • Orchiectomy
  • Prostate-Specific Antigen / metabolism
  • Prostatic Neoplasms, Castration-Resistant / genetics*
  • Prostatic Neoplasms, Castration-Resistant / pathology*
  • RNA Splicing / genetics*
  • RNA, Messenger / analysis
  • Receptors, Androgen / genetics*
  • Receptors, Androgen / physiology
  • Testosterone / pharmacology*
  • Xenograft Model Antitumor Assays

Substances

  • Androgen Receptor Antagonists
  • RNA, Messenger
  • Receptors, Androgen
  • Testosterone
  • Prostate-Specific Antigen