摘要
目的:研究维生素E琥珀酸酯(vitamin E succinate,VES)是否可通过氧化应激来诱导胃癌细胞凋亡及其可能机制。方法:分别用5、10和20μg/ml的VES处理胃癌SGC-7901细胞24h,同时设阴性对照组和甘露醇(活性氧抑制剂)预处理组。通过流式细胞技术、共聚焦技术和Western blot方法,检测了VES诱导SGC-7901细胞线粒体氧化损伤及细胞凋亡的情况,用相应的检测试剂盒检测细胞内活性氧和Ca2+含量变化情况。结果:不同浓度的VES对SGC-7901细胞均有诱导凋亡的作用。表现为随VES浓度的增加,SGC-7901细胞凋亡数量呈现增加的趋势,且SGC_7901细胞内活性氧含量及钙离子水平呈现增加的趋势,而线粒体膜电位逐渐下降。活性氧抑制剂甘露醇(1mmol/L)对VES诱导的胃癌细胞凋亡,和细胞内活性氧、钙浓度的增高及膜电位的降低均有显著抑制作用(P<0.05)。随VES浓度的增加,SGC_7901细胞内Bax蛋白的表达呈现上升的趋势,而Bcl-2蛋白的表达呈现下降趋势。甘露醇可显著降低胃癌SGC-7901细胞Bax蛋白的表达水平,同时增加Bcl-2蛋白的表达(P<0.05)。结论:VES可显著促进胃癌细胞凋亡,并且随着VES剂量的增加,VES诱导胃癌细胞凋亡的作用越强。细胞内活性氧含量增加、细胞内钙超载所导致的线粒体氧化损伤作用可能是VES诱导细胞凋亡的机制之一。
OBJECTIVE：To study the apoptosis effect of SGC-7901 cells induced by Vitamin E succinate（VES）,and explore the involvement and mechanisms of oxidative damage.METHODS：After treatment by 0,5,10,20 μg/ml VES for 24 h,methods of flow cytometry,confocal laser scanning microscopy and western blot were used to measure the mitochondrial oxidative damage and apoptosis induced by VES of SGC-7901 cells.The corresponding detection kits were used to detect the reactive oxygen species and Ca^2＋ content.At the same time,SGC-7901 cells were treated with 1 mmol/L of mannitol for 2 h,and then 20 μg/ml VES for 24 h,and the above detection methods were used.RESULTS：With the increasing concentration of VES,apoptosis showed an increasing trend.Compared with the negative control group,ΔΨm of cells in 10,20 μg/ml VES groups were significantly decreased,while the active oxygen content and the level of Ca2＋ significantly was increased.1 mmol/L of mannitol could impact the cells of oxygen and Ca2＋ contents significantly,while significantly reduce mitochondrial oxidative damage and apoptosis caused by VES.CONCLUSION：VES could significantly increase SGC-7901 apoptosis,and with increasing doses of VES,the apoptosis rate was higher.Oxidative damage caused by raised active oxygen and Ca^2＋ content increasing and ΔΨm depletion in cells may be one of the mechanisms of apoptosis induced by VES.
出处
《癌变．畸变．突变》
CAS
CSCD
2010年第3期186-190,213,共6页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金资助项目(30671757)
