该研究旨在探讨辣木籽异硫氰酸酯衍生物（MITC-21）抑制结肠癌HCT-116细胞生长和迁移的作用。采用MTT实验、克隆形成实验、流式细胞术、细胞划痕实验和蛋白印迹法，检测经不同浓度MITC-21处理的HCT-116细胞增殖、凋亡、迁移和相关蛋白表达水平的变化情况。结果表明，MITC-21呈剂量依赖性和时间依赖性的方式显著降低HCT-116细胞的存活率，半致死剂量分别为6.79 µmol/L（24 h）、3.71 µmol/L（48 h）和1.13 µmol/L（72 h）；克隆形成实验表明，MITC-21（2和4 µmol/L）显著抑制HCT-116细胞克隆形成，处理48 h后克隆形成率从100%降低到68.36%（p<0.05）和49.51%（p<0.01）；当MITC-21浓度为4 µmol/L时HCT-116发生明显的细胞形态变化；流式细胞术分析得出，HCT-116细胞的凋亡率随着MITC-21浓度和处理时间的增加而上升，处理48 h后，细胞凋亡率从14.97%增加到21.60%（p<0.001）和22.78%（p<0.001）；细胞划痕实验表明，MITC-21显著降低细胞迁移率，迁移率从61.53%降低到32.89%（p<0.01）和24.30%（p<0.001）；蛋白表达水平检测表明，MITC-21（4 µmol/L）显著增加凋亡相关蛋白Bax/Bcl-2比率（p<0.01），降低迁移相关蛋白MMP2表达水平（p<0.05）。MITC-21能够抑制HCT-116细胞增殖、诱导细胞凋亡和抑制细胞迁移。

This study aimed to investigate the inhibitory effect of the isothiocyanate derivatives from Moringa oleifera seeds (MITC-21) on the growth and migration of colon cancer HCT-116 cells. Changes in cell proliferation, apoptosis, migration and protein expression levels of HCT-116 cells treated with different concentrations of MITC-21 were determined by the MTT assay, colony formation assay, flow cytometry, wound-healing assay and western blotting. The results showed that MITC-21 treatment decreased significantly the viability of HCT-116 cells in a dose- and time-dependent manner, with the IC50 values for MITC-21 against HCT-116 cells for 24, 48 and 72 h being 6.79 µmol/L, 3.71 µmol/L, and 1.13 µmol/L, respectively. The results of the colony formation assay indicated that MITC-21 (0, 2 and 4 µmol/L) significantly decreased the colony-forming ability of HCT-116 cells and after a 48 h treatment, the colony formation rate decreased from 100% to 68.36% (p<0.05) and 49.51% (p<0.01). When MITC-21 was at 4 µmol/L, the cell morphology of HCT-116 cells changed significantly. Flow cytometry analysis showed that the apoptosis rate of HCT-116 cells increased with an increase of the dose and time of MITC-21 treatment. After a 48 h treatment, compared with that of the control cells (14.97%), the apoptosis rate of MITC-21-treated groups increased to 21.60% (p<0.001) and 22.78% (p<0.001), respectively. The wound-healing assay revealed that MITC-21 significantly inhibited the migration of HCT-116 cells, with the cell migration rate decreasing from 61.53% to 32.89% (p<0.01) and 24.30% (p<0.001), respectively. MITC-21 at 4 µmol/L significantly increased the Bax/Bcl-2 ratio and decreased the expression levels of the cell migration-related proteins MMP2. Thus, MITC-21 can inhibit the proliferation, induce apoptosis and suppress migration of HCT-116 cells.

云南省科技厅重大专项（202002AA100005）；云南省重大科技专项-绿色食品国际合作研究中心项目(2019ZG009)；云南省中青年学术和技术带头人后备人才（2018HB040）；云南省教育厅科学研究基金项目（2019J0180）