不同浓度桦木酸对人胃癌MGC-803细胞凋亡的影响

Effects of betulinic acid on apoptosis of human gastric cancer MGC-803 cells

目的:以人胃癌MCG-803细胞为实验材料,探讨不同浓度桦木酸(BA)对人胃癌MGC-803细胞凋亡的影响,为其临床应用提供依据。方法:将人胃癌MGC-803细胞分成4组,每组设置3个复孔,对照组为未加入桦木酸的MGC-803细胞,3组实验组分别加入终浓度为10、20、30μg/ml桦木酸处理细胞48 h后,通过激光共聚焦显微镜观察各组细胞形态变化;检测桦木酸对细胞Caspase-3和Caspase-9活性的影响;利用流式细胞术检测细胞线粒体膜电位变化;qRT-PCR和Western blot检测凋亡相关基因Caspase-3、Caspase-9和Cytochrome C(Cyt c)mRNA及蛋白水平的表达变化。结果:与对照组比较,各处理组Caspase-3和Caspase-9活性显著升高(P<0.01),线粒体膜电位显著下降(P<0.01),Caspase-3、Caspase-9和Cyt c mRNA及蛋白表达均显著上调(P<0.01)。结论:在终浓度为10~30μg/ml浓度范围内,桦木酸通过调节Caspase-3、Caspase-9和Cyt c的表达诱导人胃癌MGC-803细胞凋亡。

Objective:The effects of betulinic acid(BA)on apoptosis of human gastric cancer MGC-803 cells was investigated by using human gastric cancer MGC-803 cells as experimental materials,and the basis for its clinical application was provided.Methods:The human gastric cancer MGC-803 cells were divided into 4 groups,each group was set with 3 replicates.The control group was MGC-803 cells without being added betulinic acid;the other 3 groups of experimental groups were treated with betulinic acid at final concentrations of 10,20 and 30μg/ml respectively.Cells were treated with betulinic acid of different concentrations for 48 h.Laser confocal microscope was used to observe morphological changes of MGC-803.The activities of Caspase-3 and Caspase-9 were detected by an assay kit.Flow cytometry was applied to determine mitochondrial membrane potential.The mRNA and protein levels of Caspase-3,Caspase-9 and Cyt c were also detected by qRT-PCR and Western blot,respectively.Results:Compared with the control group,the activities of Caspase-3 and caspase-9 were increased(P<0.01),while the mitochondrial membrane potential was decreased significantly(P<0.01).The mRNA and protein expressions of Caspase-3,caspase-9 and Cyt c were up-regulated significantly(P<0.01).Conclusion:In the final concentration range of 10~30μg/ml,betulinic acid can induce apoptosis of human gastric cancer MGC-803 cells by regulating the expression of Caspase-3,Caspase-9 and Cyt c.