State Key Laboratory of Traditional Chinese Medicine Syndrome, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Department of General Surgery, Chongqing General Hospital, Chongqing 400013, China.
State Key Laboratory of Traditional Chinese Medicine Syndrome, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Dongguan Institute of Guangzhou University of Chinese Medicine, Dongguan, Guangdong 510006, China.
Biomed Pharmacother. 2024 Nov;180:117529. doi: 10.1016/j.biopha.2024.117529. Epub 2024 Oct 10.
Colorectal cancer (CRC) is one of the most common and fatal diseases, yet effective therapeutic drugs are lacking in clinical settings. Gingerenone A (GA) is an active compound derived from ginger, has demonstrated anti-tumor properties. However, the efficacy of GA against CRC and its primary mechanism of action remain unclear.
MTT assay and colony formation assay were employed to evaluate cell viability. Transwell assays were utilized to assess the migratory and invasive capabilities of the cells. The effects of GA on ferroptosis related proteins were analyzed using Western blot. Levels of glutathione (GSH), malondialdehyde (MDA), Fe, and 4-hydroxynonenal (4-HNE) levels were measured with a biochemical index determination kit. Cellular reactive oxygen species (ROS) were quantified using flow cytometry. CETSA, pull-down, and co-immunoprecipitation (Co-IP) assays confirmed the interactions between GA and SLC7A11, as well as the ubiquitination promoted by SLC7A11. A xenograft mouse model was employed to validate the anticancer effect of GA in vivo.
We observed that GA significantly suppressed proliferation in human CRC cells. Additionally, GA treatment inhibited the migration, invasion, and colony formation of CRC cells. Subsequently, through the use of specific inhibitors, we discovered that the suppression of CRC cells by GA was dependent on ferroptosis rather than autophagy or apoptosis. Previous research has demonstrated that GA treatment significantly triggers ferroptosis. Mechanistically, GA treatment promotes the degradation of the SLC7A11 protein, which plays a crucial role in ferroptosis. Notably, the knockdown of SLC7A11 abolished the detrimental effects of GA on the proliferation of CRC cells and reversed GA-induced ferroptosis in CRC cells both in vivo and in vitro. Further research has shown that GA can directly bind to the SLC7A11 protein and promote its ubiquitination.
Our research provides compelling evidence that GA may serve as a potential agent for suppressing the progression of CRC by inducing ferroptosis and promoting the ubiquitination and degradation of SLC7A11.
结直肠癌（CRC）是最常见和最致命的疾病之一，但在临床环境中缺乏有效的治疗药物。姜烯酮 A（GA）是一种从生姜中提取的活性化合物，具有抗肿瘤特性。然而，GA 对 CRC 的疗效及其主要作用机制尚不清楚。
采用 MTT 法和集落形成实验评估细胞活力。使用 Transwell 实验评估细胞的迁移和侵袭能力。采用 Western blot 分析 GA 对铁死亡相关蛋白的影响。使用生化指标测定试剂盒测定谷胱甘肽（GSH）、丙二醛（MDA）、Fe 和 4-羟基壬烯醛（4-HNE）的水平。使用流式细胞术定量细胞内活性氧（ROS）。CETSA、下拉和免疫共沉淀（Co-IP）实验证实了 GA 与 SLC7A11 之间的相互作用，以及 SLC7A11 促进的泛素化。采用异种移植小鼠模型在体内验证 GA 的抗癌作用。
我们发现 GA 显著抑制人 CRC 细胞的增殖。此外，GA 处理抑制 CRC 细胞的迁移、侵袭和集落形成。随后，通过使用特异性抑制剂，我们发现 GA 对 CRC 细胞的抑制作用依赖于铁死亡而不是自噬或凋亡。先前的研究表明，GA 处理显著引发铁死亡。在机制上，GA 处理促进 SLC7A11 蛋白的降解，该蛋白在铁死亡中起关键作用。值得注意的是，SLC7A11 的敲低消除了 GA 对 CRC 细胞增殖的有害影响，并在体内和体外逆转了 GA 诱导的 CRC 细胞铁死亡。进一步的研究表明，GA 可以直接与 SLC7A11 蛋白结合并促进其泛素化。
我们的研究提供了有力的证据，表明 GA 可以通过诱导铁死亡和促进 SLC7A11 的泛素化和降解来抑制 CRC 的进展，可能成为一种潜在的抑制 CRC 进展的药物。
