活性氧在2-甲氧雌二醇诱导骨尤文肉瘤细胞凋亡中的作用  

Effect of cellular reactive oxygen species on SK-N-MC Ewing sarcoma cells upon apoptosis induction by 2-Methoxyestradiol

作　　者：李承罡[1] 贺曼[2] 张聪[2] 郝素华[2] 关海山[1] 冯浩宇[1] 陈晨[1] 王春芳[3] 

机构地区：[1]山西医科大学第二医院骨科,太原030001 [2]山西省肿瘤医院预防保健科 [3]山西医科大学医学实验中心

出　　处：《肿瘤研究与临床》2008年第9期592-596,共5页Cancer Research and Clinic

摘　　要：目的研究在2-甲氧雌二醇（2-ME）诱导SK—N—MC尤文肉瘤细胞凋亡的过程中，调控细胞活性氧水平（ROS）的环节及ROS引发的下游事件。方法利用流式细胞仪、clark氧电极等手段及撤药实验、线粒体通透性转换（PT）孔干预等方法检测2-ME作用下凋亡的可逆性及呼吸链活性、线粒体跨膜电势（△ψm）、细胞ROS水平等参数的变化，并考察它们之间的关系。结果2-ME诱导SK—N—MC细胞发生依赖于ROS的细胞凋亡。2-ME作用3h内撤药，至24h无明显凋亡发生；3h后撤药，至24h有明显凋亡。2-ME作用下，线粒体呼吸链活性被抑制，ROS生成加速；△ψm经历“升高-降低”变化，3h时开始降低，PT孔稳定剂可维持△ψm不降低；ROS水平持续升高，3h时有跳跃性增高，胛孔稳定剂对ROS升高趋势无明显影响。结论2-ME作用引起sK—N—MC细胞呼吸链的抑制，引发△ψm的升高，由此造成ROS生成加速及ROS水平升高，当ROS水平突破-定阈值，引发了胛孔的不可逆开放及△ψm的消解乃至崩溃，并最终使细胞走向凋亡。ROS是2-ME诱导凋亡信号传导中的-个环节。Objective To explore the regulation of ROS level and ROS-triggered downstream events on SK-N-MC Ewing sarcoma cells upon apoptosis induction by 2-Methoxyestradiol （2-ME）. Methods To detect the reversibility of apoptosis and the alternation of activity of respiratory chain, mitochondria transmembrane potential （△ψm）, and cellular ROS level and to explore their association with flow cytometry, clark oxygen electronic node analysis, drug-removal design, and permeability transition （PT） pore stablizing agent. Results SK-N-MC cells were induced to ROS-dependent apoptosis. Apoptosis occured irreversibly after2-ME treatment for 3 h. Upon 2-ME treatment, the activity of respiratory chain was inhibited and the ROS generation was accelerated; the △ψm underwent the increasing within 3h but decreasing after 3h which could be reversed by PT pore stablizing; the ROS level underwent the continuous increasing and PT pore stablizing had no obvious effect on it. Conclusion 2-ME causes the acceleration of ROS generation via inhibiting the activity of respiratory chain and elevating the level of △ψm . ROS plays a signaling role and when total ROS accumulate to a threshold, the PT pore opening and the collapse of △ψm could be induced irreversibly and cell is eventually introduced to death.

关 键 词：活性氧 细胞凋亡 肉瘤 尤文 2-甲氧雌二醇 

分 类 号：R73[医药卫生—肿瘤]