Cuproptosis, a novel copper-induced cell death pathway, is linked to mitochondrial respiration and mediated by protein lipoylation. The discovery of cuproptosis unfolds new areas of investigation, particularly in cancers. The present study aimed to explore the role of cuproptosis in colorectal cancer progression. The genetic alterations of cuproptosis in colon cancer were evaluated using a database. MTT assays, colony formation, and flow cytometry were used to examine the effect of elesclomol-Cu and 4-Octyl itaconate (4-OI) on colorectal cancer cell and oxaliplatin-resistant cell viability. The anti-tumor effect of elesclomol with 4-OI was verified in vivo assay. The results showed that FDX1, SDHB, DLAT, and DLST genes were more highly expressed in normal tissues than those in primary tumor tissues. Patients with high expressions of these genes in tumor tissues had a better prognosis. Using MTT assay and colony formation analysis, elesclomol-Cu pulse treatment showed significant inhibition of cell viability in HCT116, LoVo, and HCT116-R cells. In addition, flow cytometry revealed elesclomol-Cu significantly promoted apoptosis. Tetrathiomolybdate, a copper chelator, markedly inhibited cuproptosis. Subsequently, we found 2-deoxy-D-glucose, a glucose metabolism inhibitor, sensitized cuproptosis. Furthermore, galactose further promoted cuproptosis. Interestingly, 4-OI significantly enhanced cuproptosis which was irrelevant to ROS production, apoptosis, necroptosis, or pyroptosis pathways. Aerobic glycolysis was inhibited by 4-OI through GAPDH, one of the key enzymes of glycolysis, sensitizing cuproptosis. Meanwhile, FDX1 knockdown weakened the ability of 4-OI to promote cuproptosis. In vivo experiments, 4-OI with elesclomol-Cu showed better anti-tumor effects. These results indicated that elesclomol-Cu rapidly halted cell growth in colorectal cancer cells and oxaliplatin-resistant cell line. Importantly, we revealed that 4-OI inhibited aerobic glycolysis by targeting GAPDH to promote cuproptosis.

Cuproptosis 是一种新型的铜诱导细胞死亡途径，与线粒体呼吸有关，并由蛋白质脂肪化介导。铜下垂的发现开启了新的研究领域，尤其是在癌症领域。本研究旨在探讨铜质下垂在结直肠癌进展中的作用。使用数据库评估了结肠癌中 cuproptosis 的遗传改变。MTT 测定、集落形成和流式细胞术用于检测艾司氯醇-铜和衣康酸 4-辛酯 (4-OI) 对结直肠癌细胞和奥沙利铂耐药细胞活力的影响。体内试验验证了 elesclomol 与 4-OI 的抗肿瘤作用。结果显示，FDX1、SDHB、DLAT、DLST基因在正常组织中的表达高于原发性肿瘤组织。这些基因在肿瘤组织中高表达的患者预后较好。使用 MTT 测定和集落形成分析，elesclomol-Cu 脉冲处理显着抑制 HCT116、LoVo 和 HCT116-R 细胞的细胞活力。此外，流式细胞术显示 elesclomol-Cu 显着促进细胞凋亡。Tetrathiomolybdate 是一种铜螯合剂，可显着抑制铜下垂。随后，我们发现 2-脱氧-D-葡萄糖，一种葡萄糖代谢抑制剂，可引起铜细胞凋亡。此外，半乳糖进一步促进了铜下垂。有趣的是，4-OI 显着增强了与 ROS 产生、细胞凋亡、坏死性凋亡或细胞焦亡途径无关的铜凋亡。有氧糖酵解被 4-OI 通过 GAPDH 抑制，GAPDH 是糖酵解的关键酶之一，使铜细胞凋亡敏感。同时，FDX1 敲低削弱了 4-OI 促进铜下垂的能力。在体内实验中，4-OI 与 elesclomol-Cu 显示出更好的抗肿瘤作用。这些结果表明，elesclomol-Cu 可迅速阻止结直肠癌细胞和奥沙利铂耐药细胞系中的细胞生长。重要的是，我们发现 4-OI 通过靶向 GAPDH 来促进铜凋亡来抑制有氧糖酵解。