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目的探讨线粒体途径在肿瘤坏死因子凋亡诱导配体(TRAIL)诱导结肠癌细胞凋亡过程中的调节作用,为临床合理用药提供理论指导。方法采用流式细胞仪技术、荧光显色技术和Western印迹技术检测TRAIL处理结肠癌细胞SW1116后,在不同时间细胞凋亡情况、线粒体完整性改变(ΔΨm、cardiolipin情况)以及线粒体下游通路细胞色素C和Caspase-9的表达情况。结果TRAIL诱发结肠癌细胞凋亡,于4 h达凋亡高峰,凋亡指数为32．98%；在4 h出现线粒体ΔΨm下降和cardiolipin丢失增加,造成其内膜损伤；细胞色素C表达及Caspase-9酶活性随时间的延长而增加,24 h酶活性达到最大峰值为(48．12±2．21)μmol·L~(-1)·h~(-1)·mg~(-1)蛋白。TRAIL诱导的线粒体损伤可被Caspase抑制剂Z-VAD．fmk所抑制。结论线粒体途径参与TRAIL诱导结肠癌细胞的凋亡过程,以Caspase依赖方式引发线粒体ΔΨm和cardiolipin丢失,造成内膜损伤,导致细胞色素C释放和Caspase-9激活,诱发凋亡。
Objective To investigate the role of mitochondrial regulation in the apoptosis of colon cancer cells induced by tumor necrosis factor apoptosis-inducing ligand (TRAIL) and provide theoretical guidance for clinical rational drug use. Methods TRAIL treatment of colon cancer cell line SW1116 was detected by flow cytometry, fluorescence colorimetry, and Western blotting. At different times, apoptosis, mitochondrial integrity (ΔΨm, cardiolipin), and downstream mitochondrial cytochrome C were detected. And Caspase-9 expression. Results TRAIL induced apoptosis of colon cancer cells, reached the peak of apoptosis at 4 h, and the apoptotic index was 32.98%. At 4 h, the decrease of mitochondrial ΔΨm and increase of cardiolipin loss caused damage of the intimal membrane; cytochrome C expression and Caspase expression. The enzyme activity of -9 increased with the prolongation of time, and the maximum peak value of enzyme activity was (48.12 ± 2.21) μmol·L -1 ·h -1 ·mg -1 for 24 h. protein. TRAIL-induced mitochondrial damage can be induced by the Caspase inhibitor Z-VAD. Fmk suppressed. Conclusion The mitochondrial pathway is involved in the apoptosis of colon cancer cells induced by TRAIL. It induces the loss of mitochondrial ΔΨm and cardiolipin in a caspase-dependent manner, causing intimal damage, leading to the release of cytochrome C and activation of Caspase-9, and induces apoptosis.