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目的:从褐环粘盖牛肝菌(Suillus luteus)子实体中分离纯化得到异牛肝菌素(iso-suillin),研究其诱导人胃癌细胞BGC-823凋亡的作用及可能的作用机制。方法:采用MTT法检测异牛肝菌素对人胃癌细胞BGC-823增殖的抑制作用;用光镜、荧光显微镜、电子显微镜和流式细胞技术检测经异牛肝菌素处理后BGC-823细胞凋亡的形态特征及凋亡率的变化;Western印迹方法检测凋亡相关基因Caspase-3、-8、-9表达的变化。结果:MTT分析表明,异牛肝菌素可明显抑制BGC-823细胞的增殖,半数抑制浓度(IC50值)为7μg/ml;形态学的观察结果:BGC-823细胞在异牛肝菌素诱导下出现典型的凋亡形态;流式细胞术检测结果显示:对照组及3.5、7、14μg/ml异牛肝菌素处理组凋亡率分别是:(0.97±0.12)%、(14.18±0.42)%、(36.29±0.64)%、(50.72±0.57)%;Western印迹显示:在异牛肝菌素的诱导下Caspase-3、-8、-9蛋白表达增高。结论:研究表明,异牛肝菌素对BGC-823细胞增殖有抑制作用;异牛肝菌素通过上调Caspase-3、-8、-9表达促进BGC-823细胞凋亡。
OBJECTIVE: To isolate and purify iso-suillin from the fruiting body of Suillus luteus, and to study its apoptosis-inducing effect on human gastric cancer cell BGC-823 and its possible mechanism. Methods: MTT assay was used to detect the inhibitory effect of isoproterenol on the proliferation of human gastric cancer cells BGC-823. The proliferation of BGC-823 cells was detected by light microscope, fluorescence microscope, electron microscope and flow cytometry The morphological changes of apoptosis and apoptosis were observed. The expression of Caspase-3, -8, -9 was detected by Western blotting. Results: MTT assay showed that isoproterenol significantly inhibited the proliferation of BGC-823 cells with the IC50 value of 7μg / ml. Morphological observation showed that when BGC-823 cells were treated with isotebepionin The results of flow cytometry showed that the apoptotic rate of the control group and 3.5, 7, 14μg / ml isoproterenol treatment group were (0.97 ± 0.12)%, (14.18 ± 0.42) ), (36.29 ± 0.64)% and (50.72 ± 0.57)%, respectively. Western blotting showed that the expression of Caspase-3, -8 and -9 increased under the induction of isoproterenol. CONCLUSION: The results showed that isoproterenol inhibited the proliferation of BGC-823 cells and isoproterenol enhanced the apoptosis of BGC-823 cells by up-regulating the expressions of Caspase-3, -8 and -9.