Rationale:
Androgen receptor splice variant 7 (AR-V7) is a leading cause of the development of
castration-resistant
prostate cancer (CRPC). However, the regulation and function of AR-V7 at levels of post-translational modifications in
prostate cancer therapy remain poorly understood. Here, we conducted a library screen of natural products to identify potential small molecules responsible for AR-V7 protein degradation in human
prostate cancer cell lines. Methods: A natural product library was used to screen the inhibitor of AR-V7. Co-IP and biomass spectrum assays were used to identify the AR-V7-interacting
proteins, whereas western blot, confocal microscopy,
RNA interfering, and gene transfection were used to validate these interactions. Cell viability, EDU staining, and colony formation assays were employed to detect cell growth and proliferation. Flowcytometry assays were used to detect the distribution of cell cycle. Mouse xenograft models were used to study the anti-CRPC effects in vivo. Results: This screen identified
rutaecarpine, one of the major components of the Chinese medicine Evodia rutaecarpa, as a novel chemical that selectively induces AR-V7 protein degradation via K48-linked ubiquitination. Mechanically, this effect relies on
rutaecarpine inducing the formation of a GRP78-AR-V7
protein complex, which further recruits the
E3 ligase SIAH2 to directly promote the ubiquitination of AR-V7. Consequently, the genetic and pharmacological activation of the GRP78-dependent AR-V7 protein degradation restores the sensitivity of
castration-resistant
prostate cancer to anti-
androgen therapy in cell culture and animal models. Conclusions: These findings not only provide a new approach for overcoming
castration-resistance in
prostate cancer therapy, but also increase our understanding about the interplay between
molecular chaperones and
ubiquitin ligase in shaping protein stability.