Li Weipeng, Li Yangyang, Lin Fengchi, Guo Huan, Zhou Haihong, Li Haining, Su Haixiang, Wang Tao
Translational Medicine Research Center, Sun Yat-sen University Cancer Center Gansu Hospital, Lanzhou, 730050, Gansu, China.
School of Basic Medical Sciences, Gansu University of Chinese Medicine, Lanzhou, 730000, Gansu, China.
Sci Rep. 2025 Jul 1;15(1):21271. doi: 10.1038/s41598-025-04415-4.
In recent years, the incidence of colorectal cancer is still on the rise. The killing of tumor cells through chemotherapy and/or radiation therapy is the mainstay of clinical anticolorectal cancer therapy, but is limited by drug and radiation resistance of tumor cells. Ferroptosis, a novel mode of programmed cell death, plays an important role in antitumor therapy. Ferroptosis inducers have been extensively studied as a strategy to target drug-resistant cancers. The aim of this study is to investigate the mechanism by which hydroxytyrosol (HT) induces ferroptosis in colorectal cancer cells via the Nrf2 signaling pathway. The goal of this study is to use network pharmacology and molecular docking approaches to screen and confirm hydroxytyrosol targets for the treatment of colorectal cancer. The response of colorectal cancer cells to hydroxytyrosol was assessed by cell viability, colony formation assay and scratch assay. Additionally, molecular techniques, including Western blotting and fluorescent probe technology, were employed. The network pharmacological screen identified 14 core targets. Among these genes, nuclear factor-erythroid 2 related factor 2 (Nrf2) was identified as the top target. Molecular docking revealed enhanced binding activity for HT with targets related to oxidative stress, including Nrf2, NAD(P)H quinone oxidoreductase 1 (NQO1), thioredoxin reductase 1 (TrxR1), prostaglandin-endoperoxide synthase 2 (PTGS2) and aldo-keto reductase 1C3 (AKR1C3). HT-induced ferroptosis elevates iron levels, lipid peroxidation (LPO) and reactive oxygen species (ROS), while decreasing glutathione (GSH) and mitochondrial membrane potential. Moreover, HT reduced the expression of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) proteins while increasing the expression of Tfr1 protein. Changes in the expression levels of these proteins led to an increase in soluble iron pools, which in turn promoted lipid peroxidation. Notably, the ferritin deposition inhibitor ferroprostatin-1 (Fer-1) significantly reversed this process. Additionally, the levels of protein expression of Nrf2 and NQO1 were reversed by two activators of Nrf2, bardoxolone (CDDO) and sulforaphane (SFN). In summary, we provide evidence that HT may induce ferroptosis in colorectal cancer cells. Mechanistically, HT induces ferroptosis via the Nrf2 signaling pathway.
近年来，结直肠癌的发病率仍在上升。通过化疗和/或放疗杀死肿瘤细胞是临床抗结直肠癌治疗的主要手段，但受到肿瘤细胞耐药性的限制。铁死亡是一种新型的程序性细胞死亡方式，在抗肿瘤治疗中发挥着重要作用。铁死亡诱导剂作为一种针对耐药癌症的策略已得到广泛研究。本研究旨在探讨羟基酪醇（HT）通过Nrf2信号通路诱导结直肠癌细胞铁死亡的机制。本研究的目的是利用网络药理学和分子对接方法筛选并确认羟基酪醇治疗结直肠癌的靶点。通过细胞活力、集落形成试验和划痕试验评估结直肠癌细胞对羟基酪醇的反应。此外，还采用了包括蛋白质印迹和荧光探针技术在内的分子技术。网络药理学筛选确定了14个核心靶点。在这些基因中，核因子红细胞2相关因子2（Nrf2）被确定为首要靶点。分子对接显示HT与氧化应激相关靶点的结合活性增强，这些靶点包括Nrf2、NAD（P）H醌氧化还原酶1（NQO1）、硫氧还蛋白还原酶1（TrxR1）、前列腺素内过氧化物合酶2（PTGS2）和醛糖还原酶1C3（AKR1C3）。HT诱导的铁死亡会提高铁水平、脂质过氧化（LPO）和活性氧（ROS），同时降低谷胱甘肽（GSH）和线粒体膜电位。此外，HT降低了溶质载体家族7成员11（SLC7A11）和谷胱甘肽过氧化物酶4（GPX4）蛋白的表达，同时增加了转铁蛋白受体1（Tfr1）蛋白的表达。这些蛋白质表达水平的变化导致可溶性铁池增加，进而促进脂质过氧化。值得注意的是，铁蛋白沉积抑制剂铁抑素-1（Fer-1）显著逆转了这一过程。此外，Nrf2的两种激活剂巴多昔芬（CDDO）和萝卜硫素（SFN）逆转了Nrf2和NQO1的蛋白表达水平。综上所述，我们提供的证据表明HT可能诱导结直肠癌细胞发生铁死亡。从机制上讲，HT通过Nrf2信号通路诱导铁死亡。
