紫杉醇诱导ASTC-a-1细胞的程序性死亡机制研究  

Real-time fluorescence analysis of taxol-induced human lung adenocarcinoma(ASTC-a-1) PCD

作　　者：陈同生[1] 王小平[2] 孙磊[1] 王会营[1] 王龙祥[1] 

机构地区：[1]华南师范大学激光生命科学研究所、激光生命科学教育部重点实验室,广州510631 [2]暨南大学附属第一医院麻醉科,广州510632

出　　处：《光电子．激光》2008年第12期1706-1709,共4页Journal of Optoelectronics·Laser

基　　金：国家自然科学基金资助项目(30670507)

摘　　要：利用质粒转染技术和显微荧光成像技术在活细胞中研究了紫杉醇诱导人肺腺癌细胞(ASTC-a-1)的程序性细胞死亡(PCD)过程,利用基因荧光探针分别标记细胞以及细胞的内质网。实验结果表明紫杉醇诱导的细胞质肿胀主要是由于内质网的肿胀引起的;利用rhodamine123标记细胞内的线粒体,研究了紫杉醇对线粒体大小的影响,结果表明紫杉醇诱导了线粒体的肿胀;caspases广谱抑制剂z-VAD-fmk对紫杉醇诱导的细胞肿胀没有影响,表明caspases并没有参与紫杉醇诱导细胞质肿胀的调控过程;Hoechst33258和PI染色实验结果表明紫杉醇没有引起细胞核的浓缩和DNA的断裂以及细胞膜的破裂。以上研究结果进一步确认紫杉醇诱导ASTC-a-1的PCD过程类似paraptosis方式,既不是传统的凋亡方式,也不是坏死方式。The characteristics of taxol-induced-ASTC-a-1 PCD resembling paraptosis were studied by fluorescence probes and confocal microscope. The erRFP plasmid was used to probe the endoplasmic reticulum, and the GFP plasmid was used to probe the change of cells shape. Rhodamine 123 was used to probe the mitochongdria, and the Hoechst33258 was used to probe the nuclear condensation and DNA cleavage, and PI was used to verify the breakage of cells membrane. Our results show that taxol induced ASTC-a-1 cell paraptosis is caused by progressive swelling of mitochondria and the endoplasmic reticulum,and the taxol-induced concentration-dependent cytoplasm vacuolization is independent of caspases inhibited by z- VAD-fmk. Hoechst33258 experimental results show that taxol did not induce nuclear condensation cell apoptosis and cells membrane breakage which is the typical characteristics of cell necrosis. Taxol induces the ASTC-a-1 cells paraptosis by cytoplasm vacuolization due to swelling of the endoplasmic reticulum and the mitochondria.

关 键 词：紫杉醇(tax01) paraptosis 细胞质空泡化 程序性细胞死亡(PCD) 

分 类 号：R318.51[医药卫生—生物医学工程]