白屈菜红碱对人结直肠癌细胞凋亡的影响及其机制研究

Effect of Chelerythrine on Apoptosis of Human Colorectal Cancer Cells and Its Mechanism

关键词:  结直肠癌  细胞凋亡  氧化应激  线粒体功能障碍

Key words:  colorectal cancer  apoptosis of cells  oxidative stress  mitochondrial dysfunction

摘要:

目的 研究白屈菜红碱（chelerythrine，CHE）对体外结直肠癌细胞的生长抑制作用，并探讨其作用机制。方法 MTT测定结肠癌细胞存活率，应用流式细胞仪检测CHE处理后活性氧（reactive oxygen species，ROS）的积累和细胞凋亡情况，JC-1荧光染料法检测细胞线粒体膜电位改变，应用荧光显微镜和Western blotting验证ROS积累诱导的线粒体功能障碍。结果 CHE对HCT-116和RKO细胞发挥剂量依赖性细胞毒作用，该作用与ROS介导的凋亡蛋白的表达有关。此外，CHE能够降低伴随线粒体功能障碍的线粒体膜电位。结论 CHE通过ROS介导的线粒体功能障碍和JNKs途径抑制CRC细胞生长并诱导细胞凋亡，提示CHE可能成为潜在的治疗CRC的候选药物。

Abstract:

OBJECTIVE To investigate the inhibitory effect of chelerythrine(CHE) on the growth of colorectal cancer cells in vitro and explore its mechanism. METHODS MTT assay was used to determine the survival rate of colon cancer cells, and flow cytometry was applied to detect reactive oxygen species(ROS) accumulation and apoptosis after CHE treatment. JC-1 fluorescent method was adopted to detect cellular mitochondria potentials while using fluorescent microscope and Western blotting method to validate that accumulation of ROS inducing mitochondrial dysfunction. RESULTS CHE had a dose-dependent cytotoxic effect on HCT-116 and RKO cells, which was related to the expression of ROS mediated apoptosis proteins. In addition, CHE could reduce the mitochondrial membrane potential associated with mitochondrial dysfunction. CONCLUSION CHE inhibits cell growth and induces CRC cell apoptosis through ROS mediated mitochondrial dysfunction and JNKs pathway, suggesting CHE may be a potential candidate drug for CRC therapy.