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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。
Objective: To study whether staurosporine can induce apoptosis in NG108-15 cells and its effect on the expression of several apoptosis-related genes. Methods: Morphological changes were observed by phase contrast microscopy, fluorescence microscopy and transmission electron microscopy. DNA ladder was detected by agarose gel electrophoresis. Western blotting was used to detect the expression of apoptosis-related genes. Results: The apoptotic morphological changes of NG108-15 cells after exposure to staurosporine at 0.1 μmol / L showed that DNA ladder appeared at 6 hours after staurosporine treatment, The expression began to rise at 6h after staurosporine treatment, reached the peak at 12h, decreased after 24h, and the expression of Bcl-2 protein significantly increased at 3h after staurosporine treatment, then decreased gradually. After staurosporine treatment At 6 h, the cleaved caspase-3 product appeared, but the expression of Cdk5 protein showed no significant change. The p53 protein expression decreased after 12 h treatment with staurosporine. CONCLUSION: Staurosporine induces the apoptosis of NG108-15 cells, which may be related to the increase of Bax protein expression and cleavage of caspase-3, but not to p53 and Cdk5.