秦皮甲素对人肺癌细胞A549凋亡影响

Impact of esculin on apoptosis of A549 lung cancer cells

目的探讨秦皮甲素对肺癌细胞A549增殖、凋亡的影响及机制。方法人肺癌细胞A549分为对照组、阳性对照组(5-Fu 0.77 mmol/L)、秦皮甲素0.4、0.8、1.6 mmol/L组,四甲基偶氮噻唑蓝比色法检测肺癌细胞A549的增殖率,流式细胞仪检测细胞周期、细胞凋亡率和线粒体膜电位的变化,免疫印迹法检测Bax,Bcl-2,Caspase-3蛋白表达改变。结果秦皮甲素可抑制人肺癌细胞A549的增殖,IC50为0.4 mmol/L;降低线粒体膜电位并诱导凋亡;与对照组比较,人肺癌细胞A549经秦皮甲素1.6 mmol/L组处理后,S期细胞比例[(20.35±0.26)%]升高,G2/M期细胞比例[(25.46±0.17)%]降低;与对照组比较,秦皮甲素1.6 mmol/L组细胞凋亡率(29.15%)明显升高,Bax和Caspase-3蛋白表达增强,Bcl-2蛋白表达受到抑制,呈剂量-效应关系。结论秦皮甲素可以通过线粒体途径诱导人肺癌细胞A549凋亡。

Objective To explore the effect of esculin on proliferation and apoptosis of A549 lung cancer cells.Methods The A549 lung cancer cells were assigned into a negative control group,a positive control group（0. 77 mmol/L of 5-fluorouracil）,and three esculin groups（0. 4,0. 8,and 1. 6 mmol/L esculin）. Methyl thiazolyl tetrazolium（M TT）method was used to observe the grow th of A549 cells. Cell cycle,cell apoptosis and mitochondrial membrane potential of the A549 lung cancer cells were detected with flow cytometry,and the expressions of Bax,Bcl-2,and caspase-3 protein were detected with Western blot. Results Esculin could inhibit the grow th of A549 lung cancer cells,with a 50% inhibitory concentration（IC50） of 0. 4 mmol/l,and reduce mitochondrial membrane potential of the cells. Compared with the control group,the proportion of cells in S phase（20. 35 ± 0. 26%） and cell apoptosis rate（29. 15%） increased,and the ratio of cells in G2/Mphase decreased（25. 46 ± 0. 17%） for the A549 lung cancer cells treated with 1. 6 mmol/L esculin. In addition,esculin could enhance the expressions of Bax and caspase-3 protein and inhibit the expression of Bcl-2protein in dose-dependent manner. Conclusion Esculin can induce apoptosis of A549 lung cancer cells through mitochondrial pathw ay.