Staurosporine induces apoptosis in NG108-15 cells  

星形孢菌素诱导NG108-15细胞凋亡(英文)

作　　者：张百芳[1] 彭芳芳[1] 章江洲[1] 武栋成[1] 

机构地区：[1]武汉大学医学院生物化学教研室

出　　处：《Acta Pharmacologica Sinica》2003年第7期663-669,725,共8页中国药理学报（英文版）

基　　金：Project supported by Hubei Health Bureau, № WJ01553.

摘　　要：AIM: To observe if staurosporine induced apoptosis in NG108-15 cells and its effect on protein expression level of several genes related to apoptosis. METHODS: Phase-contrast microscopy, fluorescence microscopy, and transmission electron microscopy were used to observe staurosporine-induced morphological changes. Agarose gel electrophoresis was used to detect DNA fragmentation. Western blots were used to measure protein expression level of several genes related to apoptosis. RESULTS: Cells treated with staurosporine 0.1 μmol/L showed typical morphological changes of apoptosis. A 'ladder' pattern representing fragmentation of DNA into oligonucleosome length fragments was observed after 6 h of staurosporine treatment and sustained until 24 h. The Bax expression increased significantly at 6 h after exposure to staurosporine, peaked at 12 h compared with vehicle cultures, and decreased at 24 h. The Bcl-2 expression increased and reached the highest level at 3 h. It was then decreased gradually but still higher than normal expression level. There was an obvious caspase-3 cleavage at 6 h after exposing the cells to staurosporine. Treatment with staurosporine for 12 h markedly decreased the expression of p53 protein. Cdk5 protein expression did not have obvious changes after the cells were exposed to staurosporine. CONCLUSION: Staurosporine induced apoptotic death in NG108-15 cells. Cells might die via a pathway that is dependent on Bax expression but independent of p53, and caspase-3 cleavage was involved.目的:研究星形孢菌素是否能引起NG108-15细胞的凋亡及它对数种与凋亡相关基因蛋白表达水平的影响。方法:用相差显微镜、荧光显微镜和透射电镜观察形态学变化;琼脂糖凝胶电泳检测DNA梯带:免疫印迹法检测凋亡相关基因蛋白的表达水平。结果:经星形孢菌素0.1μmol/L处理后NG108-15细胞呈典型的凋亡形态学变化,星形孢菌素处理后6h即出现DNA凋亡梯带,可持续至24h,Bax蛋白表达在星形孢菌素处理后6h开始上升,12h至最高峰,24h后下降,Bcl-2蛋白表达在星形孢菌素处理后3h明显上升,随后逐渐下降,星形孢菌素处理后6h可见caspase-3切割产物出现,但Cdk5蛋白的表达水平未见明显变化,p53蛋白表达在星形孢菌素处理12h后下降。结论:星形孢菌素诱导了NG108-15细胞的凋亡,可能与Bax蛋白表达的增加及caspase-3切割有关,而与p53和Cdk5无明显相关性。

关 键 词：STAUROSPORINE NG108-15 cells APOPTOSIS DNA fragmentation bax genes bcl-2 genes p53 genes cyclin-dependent kinases 

分 类 号：R96[医药卫生—药理学]