CAJ | 학술논문

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3 h内撤药,至24 h无明显凋亡发生;3 h后撤药,至24 h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,3 h时有跳跃性增高,PT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水平突破一定阈值,引发了PT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一个环节.
목적 연구재2-갑양자이순(2-ME)유도SK-N-MC우문육류세포조망적과정중,조공세포활성양수평(ROS)적배절급ROS인발적하유사건.방법 이용류식세포의、clark양전겁등수단급철약실험、선립체통투성전환(PT)공간예등방법검측2-ME작용하조망적가역성급호흡련활성、선립체과막전세(△ψm)、세포ROS수평등삼수적변화,병고찰타문지간적관계.결과 2-ME유도SK-N-MC세포발생의뢰우ROS적세포조망.2-ME작용3 h내철약,지24 h무명현조망발생;3 h후철약,지24 h유명현조망.2-ME작용하,선립체호흡련활성피억제,ROS생성가속;△ψm경력"승고-강저"변화,3h시개시강저,PT공은정제가유지△ψm불강저;ROS수평지속승고,3 h시유도약성증고,PT공은정제대ROS승고추세무명현영향.결론 2-ME작용인기SK-N-MC세포호흡련적억제,인발△ψm적승고,유차조성ROS생성가속급ROS수평승고.당ROS수평돌파일정역치,인발료PT공적불가역개방급△ψm적소해내지붕궤,병최종사세포주향조망.ROS시2-ME유도조망신호전도중적일개배절.
Objective To explore the regulation of ROS level and ROS-triggered downstream events on SK-N-MC Ewing sarcoma cells upon apoptasis induction by 2-Methoxyestradiol (2-ME). Methods To detect the reversibility of apoptosis and the alternation of activity of respiratory chain, mitechondria 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.