在臨床治療上，遭遇到高惡性度、對常見治療具抵抗性的癌症，現在已經越來越多見，進一步造成病人癌症的進展，甚或出現轉移。癌症細胞因細胞本身多樣的分子網路，複雜的代謝作用，使得只作用在單一目標或機轉的藥物，容易在後期失去效果。因此，研究潛在具多重目標的藥物、發展新的抑癌機轉，了解是否有其他非典型的調控細胞死亡形式，並最好是高惡性度或是有抗藥性癌症細胞較依賴的死亡方式，都是增加治療選項的好方法。我們藉由探討evodiamine這種具多重藥理作用的吲哚類生物鹼，在癌症細胞中的抑癌效果，來進一步研究是否有可能的、潛在臨床實用性的，未完全被發現或了解的抗癌機制。我們首先在人類和小鼠的肝癌細胞中，發現evodiamine會抑制細胞增殖，且其治療會增加WW domain-containing oxidoreductase (WWOX) 這種抑癌因子的表現。如果將癌細胞中WWOX的表現先予以小分子干擾核糖核酸控制，將會降低evodiamine的抑癌效果。更進一步，我們在人類的膀胱癌細胞中，發現evodiamine具細胞毒殺及細胞週期阻滯的效果。我們發現evodiamine治療使細胞中活性氧類增加，且證實其為脂質過氧化物，而evodiamine亦使保護細胞免於膜脂過氧化的磷脂過氧化氫酶glutathione peroxidase 4 (GPX4)表現下降。脂質過氧化物增加、GPX4下降，是細胞鐵依賴型死亡的特色之一，我們從另一角度，給予鐵螯合劑deferoxamine後再讓細胞接受evodiamine治療，發現細胞內脂質過氧化物產量下降，並有效抑制細胞死亡。若給予的是細胞凋亡抑制劑或是壞死性凋亡抑制劑，均不能讓癌細胞對evodiamine治療產生保護效果，證實evodiamine是造成膀胱癌細胞鐵依賴型死亡。在細胞和動物實驗中，我們也呈現evodiamine可抑制腫瘤成長、轉移能力，及上皮細胞間質化蛋白質的表現。以上發現可更深入了解evodiamine的抑癌能力，並進一步探究可能的抗癌機轉。

In clinical practice, we often encounter highly malignant cancers that resist conventional therapies, leading to disease progression and metastasis. Due to their complicated intracellular molecular networks and metabolic processes, cancer cells can easily develop resistance to drugs that only target single pathways or mechanisms, resulting in decreased efficacy in later stages. Therefore, it is crucial to investigate potential drugs with multiple targets and explore novel anti-cancer mechanisms, including atypical forms of regulated cell death, especially for high-grade or drug-resistant cancer cells, to expand treatment options.
In our study, we aim to investigate the anti-cancer effects of evodiamine, a multi-pharmacological indole alkaloid, and explore its applicability in previously unrecognized or understood anti-cancer mechanisms. Initially, we observe that evodiamine inhibits cell proliferation in both human and mouse liver cancer cells, and its treatment increases the expression of the tumor suppressor WW domain-containing oxidoreductase (WWOX). Furthermore, when we selectively interfere with WWOX expression using small interfering RNA in cancer cells, the anti-cancer effect of evodiamine is diminished. In human bladder cancer cells, we also reveal that evodiamine exhibits cytotoxicity and induces cell cycle arrest. We observe an increase in reactive oxygen species (ROS) upon evodiamine treatment, and the ROS are confirmed to be lipid peroxides. Moreover, evodiamine downregulates the expression of glutathione peroxidase 4 (GPX4), which protects cells from lipid peroxidation. Increased lipid peroxides and decreased GPX4 are characteristic features of ferroptosis. We administer iron chelator deferoxamine before evodiamine treatment, which decreases lipid peroxidation and effectively inhibits cell death. However, when administered apoptosis or necroptosis inhibitors, they do not confer protection against evodiamine treatment, thus confirming that evodiamine induces ferroptosis in bladder cancer cells. Moreover, we demonstrate that evodiamine can inhibit tumor growth, migrative ability, and the expression of epithelial-mesenchymal transition (EMT) proteins in vivo and in vitro. These findings provide a deeper understanding of the anti-cancer capabilities of evodiamine and offer insights into its potential mechanisms of action.

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