Li Lanlan, Lu Jianzhong, Fu Shengjun, Li Wenyan, Wang Ying, Wang Ke, Tao Yan, Liu Shanhui
Institute of Urology, Gansu Province Clinical Research Center for Urinary System Diseases, Lanzhou University Second Hospital, No. 82 Cuiyingmen, Lanzhou, 730030, Gansu, China.
Department of Gynaecology and Obstetrics, Lanzhou University Second Hospital, No. 82 Cuiyingmen, Lanzhou, 730030, Gansu, China.
Chin Med. 2025 May 26;20(1):71. doi: 10.1186/s13020-025-01130-0.
Prostate cancer is a major global health challenge, characterized by high morbidity and mortality rates. Traditional treatment options, including androgen deprivation therapy and chemotherapy, often lead to drug resistance. In recent years, natural compounds have garnered attention for their potential therapeutic effects. Evodiamine, a bioactive alkaloid from Evodia rutaecarpa, has demonstrated promising anti-cancer properties in various malignancies, including oral squamous cell carcinoma, breast, colorectal, and ovarian cancers. This study explores the efficacy of evodiamine in prostate cancer cells and investigates the mechanisms underlying evodiamine-induced cell death.
To investigate the effects of evodiamine on prostate cancer cells, various cell lines, including both castration-sensitive and castration-resistant variants, were treated with different concentrations of evodiamine for various durations. Cell viability, proliferation, invasion ability, and colony formation were assessed using the CCK8 assay, EdU assay, 3D matrigel drop invasion assay, and colony formation assay, respectively. The effects of evodiamine on apoptosis were analyzed using FACS, Hoechst staining, and Western blot. To evaluate its effects on ferroptosis, malondialdehyde (MDA) and glutathione (GSH) assay kits, as well as DCFH-DA and the lipid peroxidation sensor BODIPY 581/501 C11 fluorescent probes, were employed. The molecular mechanisms through which evodiamine regulates GPX4 protein instability were investigated using Western blot and TRIM26 ectopic expression. Additionally, a mouse xenograft model derived from DU145 cells was established to evaluate the in vivo effects of evodiamine and its molecular mechanisms, utilizing hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), and Western blot analysis.
Evodiamine significantly suppressed cell viability, proliferation, invasion, and colony formation in prostate cancer cells. Importantly, evodiamine-induced cell death in the PC3 and DU145 cell lines was independent of apoptosis pathway. Instead, evodiamine increased reactive oxygen species (ROS) production, lipid ROS levels and MDA levels, while decreasing GSH levels, indicating the induction of ferroptosis. The key role of ROS in evodiamine-induced ferroptosis was further confirmed by the partial reversal of cell death upon treatment with the ROS scavenger N-acetylcysteine (NAC). Mechanistically, evodiamine induced ferroptosis by destabilizing GPX4 protein in a TRIM26-dependent manner. Moreover, in vivo studies demonstrated that evodiamine significantly inhibited tumor growth and induced ferroptosis in tumor cells, highlighting its therapeutic potential.
This study demonstrates that evodiamine exerts potent antitumor effects against prostate cancer through inhibiting TRIM26-mediated stabilization of GPX4 protein and triggering ferroptosis. These findings suggest that evodiamine, a natural product derived from traditional Chinese medicine, could be a promising therapeutic agent for prostate cancer.
前列腺癌是一项重大的全球健康挑战，其发病率和死亡率都很高。包括雄激素剥夺疗法和化疗在内的传统治疗方案常常会导致耐药性。近年来，天然化合物因其潜在的治疗作用而受到关注。吴茱萸碱是一种从吴茱萸中提取的生物活性生物碱，在包括口腔鳞状细胞癌、乳腺癌、结直肠癌和卵巢癌在内的多种恶性肿瘤中已显示出有前景的抗癌特性。本研究探讨了吴茱萸碱对前列腺癌细胞的疗效，并研究了吴茱萸碱诱导细胞死亡的潜在机制。
为了研究吴茱萸碱对前列腺癌细胞的影响，使用不同浓度的吴茱萸碱对多种细胞系进行了不同时长的处理，这些细胞系包括去势敏感型和去势抵抗型变体。分别使用CCK8法、EdU法、3D基质胶滴侵袭试验和集落形成试验评估细胞活力、增殖、侵袭能力和集落形成。使用流式细胞术、Hoechst染色和蛋白质免疫印迹分析吴茱萸碱对细胞凋亡的影响。为了评估其对铁死亡的影响，使用了丙二醛（MDA）和谷胱甘肽（GSH）检测试剂盒，以及2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）和脂质过氧化传感器BODIPY 581/501 C11荧光探针。使用蛋白质免疫印迹和TRIM26异位表达研究了吴茱萸碱调节谷胱甘肽过氧化物酶4（GPX4）蛋白不稳定性的分子机制。此外，建立了源自DU145细胞的小鼠异种移植模型，利用苏木精和伊红（H&E）染色、免疫组织化学（IHC）和蛋白质免疫印迹分析评估吴茱萸碱的体内作用及其分子机制。
吴茱萸碱显著抑制前列腺癌细胞的活力、增殖、侵袭和集落形成。重要的是，吴茱萸碱诱导PC3和DU145细胞系中的细胞死亡与凋亡途径无关。相反，吴茱萸碱增加了活性氧（ROS）的产生、脂质ROS水平和MDA水平，同时降低了GSH水平，表明诱导了铁死亡。用ROS清除剂N-乙酰半胱氨酸（NAC）处理后细胞死亡的部分逆转进一步证实了ROS在吴茱萸碱诱导的铁死亡中的关键作用。从机制上讲，吴茱萸碱通过以TRIM26依赖的方式使GPX4蛋白不稳定来诱导铁死亡。此外，体内研究表明，吴茱萸碱显著抑制肿瘤生长并诱导肿瘤细胞发生铁死亡，突出了其治疗潜力。
本研究表明，吴茱萸碱通过抑制TRIM26介导的GPX4蛋白稳定并触发铁死亡，对前列腺癌发挥强大的抗肿瘤作用。这些发现表明，吴茱萸碱这种源自中药的天然产物可能是一种有前景的前列腺癌治疗药物。
