Wang Hong, Cui Chunli, Li Weiyi, Wu Hui, Sha Jianjun, Pan Jiahua, Xue Wei
Department of Urology, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Special Examinations, Qingdao Women and Children Hospital, Qingdao, China.
J Exp Clin Cancer Res. 2025 Jan 14;44(1):12. doi: 10.1186/s13046-025-03272-3.
Most patients with prostate cancer inevitably progress to castration-resistant prostate cancer (CRPC), at which stage chemotherapeutics like docetaxel become the first-line treatment. However, chemotherapy resistance typically develops after an initial period of therapeutic efficacy. Increasing evidence indicates that cancer stem cells confer chemotherapy resistance via exosomes. This study demonstrated that AGD1, derived from prostate cancer stem cells (PCSCs), enhanced the stemness of prostate cancer cells and reduced the therapeutic effect of docetaxel in CRPC.
Quantitative real-time PCR (qPCR) was employed to determine the expression levels of AGD1 and METTL13 mRNAs in PCSCs and exosomes. Protein expression levels were examined using western blots and dot blots. The potential functions of AGD1 and METTL13 in CRPC were investigated through cell proliferation assay, Transwell assay, EdU incorporation assays, Annexin V-FITC/PI staining, and sphere formation assays. To uncover the underlying mechanisms of AGD1, RNA pull-down assay, RIP, co-Immunoprecipitation (co-IP), mass spectrometry (MS), Methylated RNA immunoprecipitation (MeRIP) and single-base elongation and ligation-based qPCR amplification method (SELECT) were performed. The effects of AGD1 and METTL13 on CRPC development and metastasis under docetaxel treatment were analyzed using a xenograft mouse model and an organoid model. Additionally, liposomal-chitosan nanocomplex drug delivery systems were designed to explore AGD1's role in regulating docetaxel treatment resistance in CRPC.
AGD1 expression was upregulated in PCSCs and exosomes. Downregulating AGD1 enhanced the sensitivity of CRPC to docetaxel treatment by inhibiting their stemness, with the reverse also being true. RNA pull-down, combined with MS, co-IP and RIP assays, demonstrated that AGD1 binds to METTL13 and USP10, forming a complex that facilitates METTL13 protein accumulation through USP10-induced deubiquitination. MeRIP assay and SELECT assay revealed that METTL13 transcriptionally controls the mRNA decay of CD44 via m6A methylation. Additionally, this process activates the pSTAT3/PI3K-AKT signaling pathway. Organoid models and liposomal-chitosan nanocomplex drug delivery systems showed that reducing AGD1 expression enhanced the therapeutic effect of docetaxel in CRPC.
AGD1 mediates the stemness and apoptosis of PCSCs and promotes docetaxel treatment resistance by enhancing tumor growth and metastasis through USP10/METTL13-mediated CD44 mRNA decay in CRPC.
大多数前列腺癌患者不可避免地会进展为去势抵抗性前列腺癌（CRPC），在此阶段，多西他赛等化疗药物成为一线治疗方案。然而，化疗耐药性通常在初始治疗有效一段时间后出现。越来越多的证据表明，癌症干细胞通过外泌体赋予化疗耐药性。本研究表明，源自前列腺癌干细胞（PCSCs）的AGD1增强了前列腺癌细胞的干性，并降低了多西他赛在CRPC中的治疗效果。
采用定量实时PCR（qPCR）测定PCSCs和外泌体中AGD1和METTL13 mRNA的表达水平。使用蛋白质印迹和斑点印迹检测蛋白质表达水平。通过细胞增殖试验、Transwell试验、EdU掺入试验、Annexin V-FITC/PI染色和球体形成试验研究AGD1和METTL13在CRPC中的潜在功能。为了揭示AGD1的潜在机制，进行了RNA下拉试验、RIP、免疫共沉淀（co-IP）、质谱（MS）、甲基化RNA免疫沉淀（MeRIP）和基于单碱基延伸和连接的qPCR扩增方法（SELECT）。使用异种移植小鼠模型和类器官模型分析了AGD1和METTL13在多西他赛治疗下对CRPC发展和转移的影响。此外，设计了脂质体-壳聚糖纳米复合药物递送系统，以探索AGD1在调节CRPC中多西他赛治疗耐药性方面的作用。
AGD1在PCSCs和外泌体中的表达上调。下调AGD1通过抑制CRPC的干性增强了其对多西他赛治疗的敏感性，反之亦然。RNA下拉试验结合MS、co-IP和RIP试验表明，AGD1与METTL13和USP10结合，形成一个复合物，通过USP10诱导的去泛素化促进METTL13蛋白积累。MeRIP试验和SELECT试验表明，METTL13通过m6A甲基化转录控制CD44的mRNA降解。此外，这一过程激活了pSTAT3/PI3K-AKT信号通路。类器官模型和脂质体-壳聚糖纳米复合药物递送系统表明，降低AGD1表达增强了多西他赛在CRPC中的治疗效果。
AGD1介导PCSCs的干性和凋亡，并通过USP10/METTL13介导的CRPC中CD44 mRNA降解增强肿瘤生长和转移，从而促进多西他赛治疗耐药性。
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