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目的研究姬松茸多糖(PAB)对人肝癌细胞Bel-7402凋亡的影响及其机制。方法用不同浓度的PAB与小鼠脾淋巴细胞共同培养制备上清;应用流式细胞术分析各组Bel-7402细胞凋亡情况、细胞内p53蛋白的表达以及肿瘤细胞内Ca2+水平的变化;应用透射电子显微镜观察细胞超微结构变化;应用荧光显微镜观察细胞内p53蛋白表达的变化。结果与阴性对照组相比较,通过流式细胞仪检测,治疗组细胞凋亡率和细胞内Ca2+水平显著升高(P<0.01);通过流式细胞仪和荧光显微镜检测细胞内p53蛋白的表达无显著差异,透射电子显微镜观察显示出细胞凋亡的典型形态学变化。结论PAB具有诱导Bel-7402细胞凋亡的作用。
Objective To study the effect of polysaccharide from Agaricus Blazei Murill on the apoptosis of human hepatoma Bel-7402 cells and its mechanism. Methods The supernatants were prepared by co-cultivation of mouse spleen lymphocytes with different concentrations of PAB. Flow cytometry was used to analyze the apoptosis of Bel-7402 cells, the expression of intracellular p53 protein and the changes of intracellular Ca2+ levels. The transmission electron microscopy was used to observe the ultrastructure changes of the cells. The expression of p53 protein in the cells was observed by fluorescence microscope. Results Compared with the negative control group, the apoptosis rate and intracellular Ca2+ level in the treated group were significantly increased by flow cytometry (P<0.01); the expression of p53 protein in cells was detected by flow cytometry and fluorescence microscopy. No significant difference was observed by transmission electron microscopy, showing typical morphological changes of apoptosis. Conclusion PAB can induce apoptosis of Bel-7402 cells.