Evodiamine, a novel alkaloid, was isolated from the fruit of tetradium. It exerts a diversity of pharmacological effects and has been used to treat gastropathy, hypertension, and eczema. Several studies reported that evodiamine has various biological effects, including anti-nociceptive, anti-bacterial, anti-obesity, and anti-cancer activities. However, there is no research regarding its effects on drug-resistant cancer. This study aimed to investigate the effect of evodiamine on human vemurafenib-resistant melanoma cells (A375/R cells) proliferation ability and its mechanism. Cell activity was assessed using the cell counting kit-8 (CCK-8) method. Flow cytometry assay was used to assess cell apoptosis and cell cycle. A xenograft model was used to analyze the inhibitory effects of evodiamine on tumor growth. Bioinformatics analyses, network pharmacology, and molecular docking were used to explore the potential mechanism of evodiamine in vemurafenib-resistant melanoma. RT-qPCR and Western blotting were performed to reveal the molecular mechanism. The alkaloid extract of the fruit of tetradium, evodiamine showed the strongest tumor inhibitory effect on vemurafenib-resistant melanoma cells compared to treatment with vemurafenib alone. Evodiamine inhibited vemurafenib-resistant melanoma cell growth, proliferation, and induced apoptosis, conforming to a dose–effect relationship and time–effect relationship. Results from network pharmacology and molecular docking suggested that evodiamine might interact with IRS4 to suppress growth of human vemurafenib-resistant melanoma cells. Interestingly, evodiamine suppressed IRS4 expression and then inhibited PI3K/AKT signaling pathway, and thus had the therapeutic action on vemurafenib-resistant melanoma.

吴茱萸碱是从吴茱萸果实中分离出来的一种新型生物碱。它具有多种药理作用，已用于治疗胃病、高血压和湿疹。多项研究表明吴茱萸碱具有多种生物效应，包括抗伤害、抗菌、抗肥胖和抗癌活性。然而，尚无关于其对耐药癌症的影响的研究。本研究旨在探讨吴茱萸碱对人维莫非尼耐药黑色素瘤细胞（A375/R细胞）增殖能力的影响及其机制。使用细胞计数试剂盒-8 (CCK-8) 方法评估细胞活性。流式细胞术用于评估细胞凋亡和细胞周期。采用异种移植模型分析吴茱萸碱对肿瘤生长的抑制作用。利用生物信息学分析、网络药理学和分子对接来探索吴茱萸碱治疗维莫非尼耐药黑色素瘤的潜在机制。 RT-qPCR 和 Western blotting 揭示了分子机制。与单独使用威罗非尼治疗相比，吴茱萸果实的生物碱提取物对威罗非尼耐药的黑色素瘤细胞显示出最强的肿瘤抑制作用。吴茱萸碱抑制维莫非尼耐药黑色素瘤细胞的生长、增殖并诱导细胞凋亡，符合量效关系和时效关系。网络药理学和分子对接结果表明，吴茱萸碱可能与 IRS4 相互作用，抑制人类维莫非尼耐药黑色素瘤细胞的生长。有趣的是，吴茱萸碱抑制IRS4表达，进而抑制PI3K/AKT信号通路，从而对维莫非尼耐药的黑色素瘤具有治疗作用。