Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Department of Urology, Key Laboratory of Biological Targeting Diagnosis, Therapy and Rehabilitation of Guangdong Higher Education Institutes, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
J Cell Mol Med. 2024 Nov;28(22):e70210. doi: 10.1111/jcmm.70210.
Clinically, most prostate cancer (PCa) patients inevitably progress to castration-resistant prostate cancer (CRPC) with poor prognosis after androgen deprivation therapy (ADT), including abiraterone, the drug of choice for ADT. Therefore, it is necessary to explore the resistance mechanism of abiraterone in depth. Genome-wide CRISPR/Cas9 knockout technology was used to screen CRPC cell line 22Rv1 for abiraterone-resistant genes. Combined with bioinformatics, a key gene with high expression and poor prognosis in CRPC patients was screened. Then, the effects of target gene on abiraterone-resistant 22Rv1 cell function were explored by silencing and overexpression. Further, a natural product with potential targeting effect was identified and validated by molecular docking and protein expression. Molecular dynamics simulations revealed potential mechanism for the natural product affecting target protein expression. Finally, the combined anti-CRPC effects of the natural product and abiraterone were validated by cellular and in vivo experiments. Five common resistance genes (KCNJ3, COL2A1, PPP1CA, MDH2 and EXOSC5) were identified successfully, among which high PPP1CA expression had the worst prognosis for disease-free survival. Moreover, PPP1CA was highly expressed in abiraterone-resistant 22Rv1 cells. Silencing PPP1CA increased cell sensitivity to abiraterone while promoting apoptosis and inhibiting clone formation. Overexpressing PPP1CA exerted the opposite effects. Molecular docking revealed the binding mode of the natural product nodularin-R to PPP1CA with a dose-dependent manner for inhibition. Mechanistically, nodularin-R attenuates the interaction between PPP1CA and USP11 (deubiquitinating enzyme), potentially promoting PPP1CA degradation. Additionally, combination of 2.72 μM nodularin-R and 54.5 μM abiraterone synergistically inhibited the resistant 22Rv1 cell function. In vivo experiments also revealed that combination therapy significantly inhibited tumour growth and reduced inducible expression of PPP1CA. PPP1CA is a key driver for abiraterone resistance, and nodularin-R enhances the anti-CRPC effects of abiraterone by inhibiting PPP1CA.
临床上，大多数前列腺癌（PCa）患者在雄激素剥夺治疗（ADT）后不可避免地进展为去势抵抗性前列腺癌（CRPC），预后不良，包括阿比特龙，这是 ADT 的首选药物。因此，有必要深入探讨阿比特龙耐药的机制。利用全基因组 CRISPR/Cas9 敲除技术筛选阿比特龙耐药基因的 CRPC 细胞系 22Rv1。结合生物信息学，筛选出 CRPC 患者中高表达且预后不良的关键基因。然后，通过沉默和过表达研究靶基因对阿比特龙耐药 22Rv1 细胞功能的影响。进一步通过分子对接和蛋白表达鉴定并验证具有潜在靶向作用的天然产物。分子动力学模拟揭示了天然产物影响靶蛋白表达的潜在机制。最后，通过细胞和体内实验验证了天然产物和阿比特龙联合抗 CRPC 的效果。成功鉴定出 5 个常见的耐药基因（KCNJ3、COL2A1、PPP1CA、MDH2 和 EXOSC5），其中 PPP1CA 高表达对无病生存的预后最差。此外，PPP1CA 在阿比特龙耐药的 22Rv1 细胞中高表达。沉默 PPP1CA 增加了细胞对阿比特龙的敏感性，同时促进了细胞凋亡和抑制了克隆形成。过表达 PPP1CA 则产生相反的效果。分子对接揭示了天然产物 nodularin-R 与 PPP1CA 的结合模式呈剂量依赖性抑制。从机制上讲，nodularin-R 减弱了 PPP1CA 与 USP11（去泛素化酶）的相互作用，可能促进了 PPP1CA 的降解。此外，2.72 μM 的 nodularin-R 和 54.5 μM 的阿比特龙联合抑制耐药 22Rv1 细胞的功能。体内实验也表明，联合治疗显著抑制了肿瘤生长，降低了 PPP1CA 的诱导表达。PPP1CA 是阿比特龙耐药的关键驱动因素，nodularin-R 通过抑制 PPP1CA 增强了阿比特龙的抗 CRPC 作用。
