Zhang Qiao, Xu Zhijie, Liu Wanying, Cheng Zhidong, Ding Yating, Xie Yafeng, Yan Shengyu
Department of Proctology, the Second Affiliated Hospital, University of South China, Hengyang, 421001, Hunan, China; Hengyang Medical School, University of South China, Hengyang, 421001, Hunan, China.
Tissue Cell. 2025 Aug;95:102793. doi: 10.1016/j.tice.2025.102793. Epub 2025 Feb 28.
Gastrodin, an important component of traditional Chinese medicine, is gaining interest because of its anti-tumor effects. Ferroptosis is a new mode of cell death, which has emerged as a promising target for colorectal cancer (CRC) treatment. This research investigates the action mechanism of gastrodin on the process of CRC by inducing ferroptosis.
The mRNA and protein levels were measured via qRT-PCR and western blot. Cell viability was assessed by CCK-8 assay. The cell proliferation was examined using colony formation assay. Live-Dead cell staining was evaluated by Calcein-AM/PI staining. The effect of ferroptosis was evaluated by detecting the levels of reactive oxygen species (ROS), intracellular total iron, ferrous iron (Fe), malondialdehyde (MDA), glutathione (GSH) by kits, as well as the expressions of subunit solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), ferritin light chain (FTL) and acyl-CoA synthetase long chain family member 4 (ACSL4) by western blot. Co-immunoprecipitation (Co-IP) assay was applied to analyze the binding relationship between S-phase kinase-associated protein 2 (SKP2) and nuclear receptor coactivator 4 (NCOA4).
Gastrodin could induce ferroptosis in CRC cells. SKP2 ameliorated gastrodin induced ferroptosis in CRC cells. Besides, SKP2 mediated NCOA4 degradation by ubiquitination. SKP2 was involved in ferroptosis of CRC cells by regulating NCOA4. Gastrodin induced ferroptosis in CRC cells via SKP2/NCOA4 axis.
Gastrodin repressed SKP2 expression, deactivated NCOA4 ubiquitination thus elevated NCOA4 expression, and promoted ferroptosis in CRC cells.
天麻素是中药的重要成分，因其抗肿瘤作用而受到关注。铁死亡是一种新的细胞死亡模式，已成为结直肠癌（CRC）治疗的一个有前景的靶点。本研究通过诱导铁死亡来探究天麻素在CRC进程中的作用机制。
通过qRT-PCR和蛋白质印迹法检测mRNA和蛋白质水平。采用CCK-8法评估细胞活力。使用集落形成试验检测细胞增殖。通过钙黄绿素-AM/PI染色评估活死细胞染色。通过试剂盒检测活性氧（ROS）、细胞内总铁、亚铁离子（Fe）、丙二醛（MDA）、谷胱甘肽（GSH）水平，以及通过蛋白质印迹法检测溶质载体家族7成员11（SLC7A11）、谷胱甘肽过氧化物酶4（GPX4）、铁蛋白轻链（FTL）和酰基辅酶A合成酶长链家族成员4（ACSL4）的表达，以评估铁死亡的影响。应用免疫共沉淀（Co-IP）试验分析S期激酶相关蛋白2（SKP2）与核受体辅激活因子4（NCOA4）之间的结合关系。
天麻素可诱导CRC细胞发生铁死亡。SKP2可改善天麻素诱导的CRC细胞铁死亡。此外，SKP2通过泛素化介导NCOA4降解。SKP2通过调节NCOA4参与CRC细胞的铁死亡。天麻素通过SKP2/NCOA4轴诱导CRC细胞发生铁死亡。
天麻素抑制SKP2表达，使NCOA4去泛素化从而提高NCOA4表达，并促进CRC细胞的铁死亡。
