Cao Jun-Feng, Hang Kuan, Zhang Hao, Wu Zuowei, Guo Ziheng, Men Jie, Tian Jin, Li Kezhou
College of Medicine, Southwest Jiaotong University, Chengdu, 610031, Sichuan, China.
Division of Pancreatic Surgery, Department of General Surgery, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
Eur J Pharmacol. 2025 Sep 15;1003:177964. doi: 10.1016/j.ejphar.2025.177964. Epub 2025 Jul 17.
Colorectal cancer is a highly aggressive malignancy characterized by complex tumor micro-environments and significant drug resistance. We highlighted the critical role of curcumin in inhibiting tumor growth and migration, inducing cuproptosis and oxidative stress within colorectal cancer cells.
The RKO human colorectal cancer cell line was utilized to conduct various in vitro assays, including CCK-8, flow cytometry, scratch assays, immunofluorescence and morphological assessments using both 2D and 3D culture models to evaluate effects of curcumin on cell proliferation, apoptosis and migration. Differential gene expression analysis was performed using GSE41657 dataset and confirmed through machine learning and bioinformatics analysis. RT-qPCR, survival analysis, pan-tumor expression assessment and immune infiltration analysis were also conducted on the identified key gene targets. Computer simulations were performed to evaluate the binding stability of curcumin with cuproptosis-associated proteins.
Cellular experiments demonstrated that curcumin at 30 μg/mL significantly inhibited the proliferation and migratory capacity of 3D colorectal cancer cell spheres while promoting cellular oxidative stress and interfering with mitochondrial activity. Bioinformatics and machine learning analyses identified CALCOCO2 and HSPD1 as key targets. RT-qPCR results and immunofluorescence results confirmed that curcumin significantly downregulated the expression of CALCOCO2 and HSPD1. Molecular dynamics simulations revealed binding free energy of HSPD1/Curcumin and CALCOCO2/Curcumin were -20.25 ± 1.57 kcal/mol and -16.67 ± 2.06 kcal/mol.
This study revealed that curcumin reduced the invasiveness of colorectal cancer by downregulating CALCOCO2 and HSPD1 expression, thereby modulating the tumor microenvironment. Moreover, curcumin promoted programmed cell death in colorectal cancer through oxidative stress-induced cuproptosis.
结直肠癌是一种侵袭性很强的恶性肿瘤，其特征是肿瘤微环境复杂且具有显著的耐药性。我们强调了姜黄素在抑制结直肠癌细胞生长和迁移、诱导铜死亡和氧化应激方面的关键作用。
利用RKO人结直肠癌细胞系进行各种体外试验，包括CCK-8、流式细胞术、划痕试验、免疫荧光以及使用二维和三维培养模型进行形态学评估，以评估姜黄素对细胞增殖、凋亡和迁移的影响。使用GSE41657数据集进行差异基因表达分析，并通过机器学习和生物信息学分析进行确认。还对鉴定出的关键基因靶点进行了RT-qPCR、生存分析、泛肿瘤表达评估和免疫浸润分析。进行计算机模拟以评估姜黄素与铜死亡相关蛋白的结合稳定性。
细胞实验表明，30μg/mL的姜黄素显著抑制三维结直肠癌细胞球的增殖和迁移能力，同时促进细胞氧化应激并干扰线粒体活性。生物信息学和机器学习分析确定CALCOCO2和HSPD1为关键靶点。RT-qPCR结果和免疫荧光结果证实，姜黄素显著下调CALCOCO2和HSPD1的表达。分子动力学模拟显示，HSPD1/姜黄素和CALCOCO2/姜黄素的结合自由能分别为-20.25±1.57kcal/mol和-16.67±2.06kcal/mol。
本研究表明，姜黄素通过下调CALCOCO2和HSPD1的表达降低了结直肠癌的侵袭性，从而调节肿瘤微环境。此外，姜黄素通过氧化应激诱导的铜死亡促进结直肠癌中的程序性细胞死亡。
