Targeting GRP78-dependent AR-V7 protein degradation overcomes castration-resistance in prostate cancer therapy
| Autor: | Zhenlong Shao, Hongbiao Huang, Jinchan He, Jinbao Liu, Daolin Tang, Xiaohong Xia, Yuan Liu, Chuyi Huang, Lili Jiang, Yuning Liao |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
GRP78
Male 0301 basic medicine Ubiquitin-Protein Ligases Medicine (miscellaneous) SIAH2 Protein degradation Mice 03 medical and health sciences Prostate cancer Alkaloids 0302 clinical medicine Cell Line Tumor Androgen Receptor Antagonists medicine Animals Humans Protein Isoforms CRPC Castration Endoplasmic Reticulum Chaperone BiP Pharmacology Toxicology and Pharmaceutics (miscellaneous) Heat-Shock Proteins Mice Inbred BALB C biology Chemistry Cell growth Ubiquitination Nuclear Proteins Rutaecarpine Transfection Cell cycle medicine.disease Antineoplastic Agents Phytogenic Xenograft Model Antitumor Assays Ubiquitin ligase rutaecarpine Prostatic Neoplasms Castration-Resistant HEK293 Cells 030104 developmental biology Receptors Androgen 030220 oncology & carcinogenesis Proteolysis biology.protein Cancer research AR-V7 Research Paper |
| Zdroj: | Theranostics |
| ISSN: | 1838-7640 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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