紫杉醇诱导不依赖于Caspase-3细胞类似Paraptosis的荧光光谱分析  

Fluorescence Analysis of Taxol-Induced Paraptosis-Like Independent of Caspase-3 Activation

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,暨激光生命科学教育部重点实验室,广东广州510631 [2]暨南大学附属第一医院麻醉科,广东广州510632

出　　处：《光谱学与光谱分析》2008年第11期2623-2627,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(30670507,60378043,30470494);中国博士后科学基金项目(2005037169)资助

摘　　要：研究了紫杉醇(Taxol)诱导人肺腺癌细胞(ASTC-a-1)类似Paraptosis的特征和机理。采用CCK-8技术检测了抑制细胞活性的特性,结果表明大于70μmol.L-1浓度的紫杉醇可以显著抑制细胞活性;采用激光共聚焦扫描荧光成像从形态上检测了紫杉醇诱导细胞死亡的形态特征,表明紫杉醇诱导了类似副凋亡(Paraptosis)特征的细胞肿胀和细胞质空泡化,而且没有细胞膜皱褶、细胞核浓缩等细胞凋亡的特征;通过基因质粒转染在细胞转染稳定表达基于荧光共振能量转移(FRET)质粒SCAT3,并利用FRET受体光漂白技术和荧光光谱检测分析技术研究了紫杉醇诱导细胞类似Paraptosis过程中Caspase-3的活化特性,结果表明在紫杉醇诱导细胞质肿胀和细胞死亡的过程中Caspase-3没有被活化。以上研究结果表明紫杉醇可以通过类似Paraptosis的方式明显诱导细胞程序性死亡,该过程不依赖于Caspase-3的活性。In the present report,the authors for the first time described the characteristics of taxol-induced paraptosis-like for the human lung adenocarcinoma cells（ASTC-a1）.CCK-8 was used to assay the inhibition of taxol on the cells viability.Cell viability was inhibited obviously 24 h after taxol treatment.Confocal fluorescence scanning microscope was used to monitor the morphology changes in cells with taxol treatment.Fluorescence resonance energy transfer（FRET） and acceptor photobleaching techniques were used to analyze the caspase-3 activation in the taxol-induced cell swelling and cell dearth.Taxol induced cell swelling,cytoplasmatic vacuolization and cell death without cell shrinkage,an apoptotic feature,and membrane rupture,a necrotic feature.The emission spectra of scat3 inside living cells expressed stably with scat3 were the same for control（without taxol）.Further analysis with FRET and acceptor photobleaching techniques showed that the caspase-3 was not activated by taxol for the cytoplastic vacuoliazation cells expressed stably with scat3 plasmid,suggesting that caspase-3 is not involved in the taxol-inducecd cell swelling,cytoplasmatic vacuolization and cell death.These results show that taxol can induce a novel nonapoptotic PCD resembling the paraptosis in ASTC-a-1 cells.

关 键 词：紫杉醇(Taxol) Paraptosis 荧光光谱 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]