莱菔子素诱导结肠癌细胞株Caco-2凋亡的研究

Apoptosis in human colon carcinoma Caco-2 cells induced by sulforaphane

目的:探讨莱菔子素(SFN)诱导结肠癌细胞凋亡的抗肿瘤机制。方法:体外培养人结肠腺癌细胞Caco-2,观察10-100μmol/L SFN对Caco-2细胞增殖动力学的影响。MTT法检测细胞生长抑制率, 倒置相差显微镜及扫描电镜观察凋亡细胞形态学变化,流式细胞仪检测细胞凋亡率及细胞周期。结果: 30-100μmol/L浓度范围的SFN抑制细胞增殖,且呈剂量和时间依赖效应。倒置相差显微镜及扫描电镜观察,25μmol/L处理组无明显改变,50、75、100μmol/L处理组细胞形态学发生明显变化,典型者出现凋亡小体。流式细胞仪检测结果表明,随着SFN浓度的增高,细胞凋亡率增加,细胞周期阻滞于G0-G1期, 与对照组差异有统计学意义(P<0.01)。结论:SFN对Caco-2细胞的生长增殖具有抑制作用,呈剂量和时间依赖效应;大剂量SFN诱导Caco-2细胞凋亡。SFN对Caco-2的生长增殖抑制作用可能是通过诱导细胞凋亡、抑制细胞增殖两条途径实现的。

Objective: To investigate the apoptosis of Caco-2 cells induced by sulforaphane and to elucidate the antineoplastic mechanism in colon cancer prevention. Methods: Human colon carcinoma cell lines Caco-2 were cultured in vitro, and the antiproliferative effect of 10-100μmol/L sulforaphane was examined by means of MTT assay. Morphological variations of apoptotic cells were observed with invert phase-contrast microscope and scanning electron microscope. Meanwhile, cell cycles and apoptosis rates were evaluated by flow cytometry. Results: The cell growth was inhibited by 30-100μmol/L sulforaphane. The obvious morphological changes of cells treated with 50-100μmol/ L sulforaphane were observed with invert phase-contrast microscope, and typical apoptotic changes with scanning electron microscope. Flow cytometry analysis showed that the apoptotic ratios were increased and G0-G1 phase accumulated with concentration enhancement in sulforaphane-treated groups. Conclusion: Sulforaphane inhibits the growth of human colon carcinoma Caco-2 cells in a time- and dose-dependent manner, and sulforaphane of high concentrations can induce the apoptosis of Caco-2 cells.