大花紫玉盘素诱导肿瘤多药抗药性细胞凋亡及其机制  

Induction of apoptosis of tumor multidrug resistance cell by uvarigrin and its mechanism

作　　者：李艳芳[1] 梁永钜[1] 石智[1] 陈黎明[1] 丁岩[1] 符立梧[1] 

机构地区：[1]中山大学肿瘤防治中心

出　　处：《药学学报》2006年第3期252-256,共5页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30371659);广东省自然科学基金资助项目(2003C30313).

摘　　要：目的比较研究番荔枝内酯大花紫玉盘素(uvarigrin)诱导多药抗药性KBv200细胞及其亲本KB细胞凋亡及其机制。方法以MTT法进行细胞毒测定;用Annexin V FITC染色及流式细胞仪检测细胞凋亡。活性氧(ROS)测定以DCFH-DA标记,细胞线粒体跨膜电位(ΔΨm)测定用D iOC6标记,均以流式细胞仪检测。Caspase-9激活的测定用W estern b lotting法。结果大花紫玉盘素对KBv200细胞及其亲本KB细胞的生长均有明显的抑制作用;大花紫玉盘素不仅能介导KB细胞凋亡,而且也能介导KBv200细胞凋亡;大花紫玉盘素作用于KBv200细胞及其亲本KB细胞12,24和48 h,均引起ROS升高以及ΔΨm降低,而且呈时间依赖性。W estern b lotting方法分析显示Caspase-9被激活。结论大花紫玉盘素可能通过线粒体通路诱导细胞凋亡。Aim To study the effect of uvarigrin on mitoehondrial dependent pathway apoptosis induced by it in MDR KBv200 cells and their parental sensitive KB cells. Methods during the MTT assay was used to detect the cytotoxic effect of uvarigrin on KBv200 and KB cells. Annexin V FITC staining identified uvarigrin-induced apoptosis in KBv200 and KB ceils. These ceils underwent incubation with DCFH-DA, or DiOC6, followed by flowcytometry for the measurement of reactive oxygen species （ROS） and mitochondrial membrane potential （△ψm）, respectively. The Western blotting analysis was performed on Caspase-9 activation. Results Uvarigrin inhibited the growth of KBv200 ceils and KB ceils in vitro. Most of the uvarigrin-induced cells death was found to be due to apoptosis, as determined by Annexin V FITC staining. During the apoptosis, the level of ROS increased while the level of △ψm decreased in a time-dependent manner. Uvarigrin triggered Caspase-9 activation. Conclusion Uvarigrin induced apoptosis in KBv200 cells and KB cells probably through a mitochondria-dependent pathway.

关 键 词：大花紫玉盘素 线粒体跨膜电位 多药抗药性 细胞凋亡 

分 类 号：R963[医药卫生—微生物与生化药学] R979.1[医药卫生—药理学]