消癌平诱导caspase-3活化动态过程的实时监测  

Monitoring the Dynamics of Caspase-3 Activation Induced by Xiaoaiping

作　　者：陈同生[1] 王龙祥[1] 孙磊[1] 王会营[1] 邢达[1] 

机构地区：[1]华南师范大学激光生命科学研究所暨激光生命科学教育部重点实验室,广东广州510631

出　　处：《激光生物学报》2007年第5期532-536,共5页Acta Laser Biology Sinica

基　　金：国家自然科学基金资助项目(306705076037804330470494);中国博士后科学基金项目(2005037169)

摘　　要：消癌平是从萝摩科(Asclepiadaceae)植物通关藤(Marsdenia tenacissima)根部提取的药物,已经广泛应用于癌症和多种炎症的临床治疗,并且具有较好的疗效。将消癌平注射液与细胞培养液按照1∶1的比例混合后共同培养人肺腺癌(ASTC-a-1)细胞,通过CCK-8方法检测发现消癌平能够显著抑制细胞的增长。然后利用荧光共聚焦扫描显微镜和荧光共振能量转移(fluorescence resonance energy transfer,FRET)技术在稳定表达了SCAT3质粒的单个ASTC-a-1细胞中实时动态监测消癌平处理后SCAT3的空间分布以及SCAT3被caspase-3切割的动态过程。实验结果表明:消癌平处理后20 min中SCAT3开始向细胞核以及细胞膜部位转移,约在100min左右SCAT3被迅速切割。Xiaoaiping （XAP） isolated from the stems of Marsdenia tenacissima, an Asclepiadaceae plant, has been documented to possess functions against inflammation and cancers. In this paper, we used the CCK-8 to measure the cell activity after XAP treatment according to the 1 ： 1 of XAP and DMEM, and we found that XAP inhibited obviously the cell activity. We used confocal fluorescence microscope and fluorescence resonance energy transfer （FRET） to measure the dynamic temporal-spatial distribution of SCAT3 and caspase-3 activation in single living human lung adenocarcinoma line （ ASTC-a-1 ） cell expressing stably with the SCAT3. Experimental results showed that SCAT3 translocated from cytoplasm to nuclear, cell membrane and intracellular membrane area after 20 min of XAP treatment, and the caspase-3 was activated rapidly after about 100 min of XAP treatment.

关 键 词：消癌平 CCK-8 caspae-3 荧光共振能量转移(FRET) 激光共聚焦荧光显微镜 

分 类 号：Q631[生物学—生物物理学] Q-336