喉癌细胞线粒体去极化和凋亡研究

目的线粒体膜电位的缺失是线粒体凋亡途径的早期事件。我们用线粒体膜电位去极化剂(valinomycin)处理3个喉癌细胞系,研究它们的凋亡反应和细胞色素C释放的能力。方法 3个喉癌细胞系和对照细胞系HL-60经过valinomycin处理后,用细胞膜电位探针JC-1染色和凋亡特异性探针annexinV染色,用流式细胞仪分别测量它们的线粒体膜电位和凋亡率。Western印记用来检测细胞浆内的细胞色素C的释放。结果在经过24小时的处理后所有细胞均显示完全的线粒体去极化,只有对照细胞HL-60出现凋亡和细胞色素C释放,而喉癌细胞系没有出现。结论喉癌细胞系表现出线粒体膜电位去极化诱导的凋亡途径的缺陷,线粒体膜电位去极化而没有细胞色素C的释放。线粒体凋亡途径的缺陷是喉癌的一个特点。

Study on mitochondrial depolarization and apoptosis of laryngeal carcinoma

Objective Loss of mitochondrial membrane potential（MMP） is an early initiating event in the mitochondrial apoptotic pathway.We treat 3 laryngeal carcinoma cell lines with a mitochondrial-membrane-depolarizing agent,valinomycin and investigated the ability of depolarization to induce release of cytochrome c in these cell lines.Methods laryngeal cell and a leukemia control cell line HL60 were assayed for MMP after valinomycin treatment by staining with mitochondrial-membrane-potential-specific probe JC-1 and stained with apoptosis-specific probe annexin-V to measure their rate of apoptosis by FACS.Western blotting was also applied to detect cytoplasmic cytochrome C release.Results All cell lines demonstrated complete mitochondrial depolarization after 24 h＇s treatment.Only HL60 displayed cytochrome C release and underwent apoptosis,laryngeal cancer cell lines did not.Conclusion laryngeal carcinoma cell lines exhibit defects in mitochondrial-membrane-depolarization-induced apoptosis and no release of cytochrome C.Defects of mitochondrial apoptosis pathway are a feature of laryngeal carcinoma.