C_2-神经酰胺所致HT-29细胞凋亡中线粒体的变化

Mitochondrial changes in apoptosis of HT-29 cells induced by C_2-ceramide

目的探讨外源性C2-神经酰胺所致HT-29细胞凋亡中线粒体的变化。方法C2-神经酰胺作用人结肠癌HT-29细胞,采用透射电镜观察细胞线粒体超微结构的变化;采用Mitochondrial Apoptosis Detection Kit和流式细胞仪检测细胞凋亡时线粒体损伤及其膜电位的变化。结果C2-神经酰胺作用HT-29细胞24h,线粒体发生空泡化、变性、嵴肿胀或消失等形态学改变;此时荧光显微镜下可观察到线粒体聚集,发出红色荧光,而正常细胞线粒体发出绿色荧光。C2-神经酰胺作用细胞6h,线粒体膜电位即开始降低,呈时间-剂量-效应关系;结论线粒体参与C2-神经酰胺诱导HT-29细胞凋亡作用。

Objective To investigate the mitochondrial changes in apoptosis of HT-29 cells induced by exogenous C2-ceramide.Methods Human colon carcinoma HT-29 cells were treated with C2-ceramide.Transmission electron microscope was used to observe the the changes of mitochondrial ultramicrostucture.Mitochondrial Apoptosis Detection Kit and flow cytometry were used to observe the mitochondrial damage and the change of mitochondrial membrane potential.Results There were the morphological changes of mitochondria,such as vacuolization,degeneration and cristae swelling or disappearance in HT-29 cells treated by C2-ceramide for 24 hours.At the same time,under the fluorescence microscope,mitochondria of apoptotic cells aggregated and emitted red fluorescence,however,mitochondria of normal cells emitted green fluorescence.The mitochondrial membrane potential began the decrease after 6 hours in HT-29 cells treated with C2-ceramide in time and dose dependent manner.Conclusion Mitochondria could participate in the apoptosis of HT-29 cells induced by C2-ceramide.