莪术醇诱导胶质瘤U87细胞凋亡的实验研究

Curcumol Inducing the Apoptosis of Human Glioma Cell Line U87 Cells

目的研究莪术醇对胶质瘤U87细胞凋亡的作用,探讨其诱导U87细胞凋亡的机制。方法分别用浓度为100、200、400μM的莪术醇作用于人胶质瘤U87细胞,应用流式细胞术Annexin V/PI双染法检测细胞凋亡水平,JC-1法检测细胞线粒体膜电位变化,Caspase-3蛋白表达水平。Western blot检测p-AKT蛋白的表达水平。结果不同浓度莪术醇作用U87细胞48h后细胞凋亡率均高于对照组(F=1774.599,P<0.05);伴随着莪术醇作用浓度的增加,U87细胞凋亡率逐渐增高;与对照组相比,莪术醇使U87细胞线粒体膜电位呈不同程度降低(P<0.05),并能显著提高U87细胞中Caspase-3蛋白表达水平(P<0.05),U87细胞经莪术醇作用48h后p-AKT蛋白的表达降低(P<0.05)。结论莪术醇具有诱导人胶质瘤细胞U87凋亡作用,其机制可能是通过线粒体途径激活Caspase家族,下调p-AKT蛋白表达有关。

Objective To explore the apoptotic mechanism of human glioma U87 cells induced by curcumol( Cur). Methods After U87 cells was treated by 100,200,400μM curcumol for 48 h,the cell apoptosis rate was analyzed by Annexin V / PI staining. The mitochondrial membrane potential was measured by JC-1staining and the expression level of caspase-3 protein was detected by flow cytometry. The expression level of p-AKT protein was detected by Western blot method. Results After treatment with different doses of Cur,U87 cells showed apoptotic feature to some extent in a dose-dependent manner,and there was statistical significance compared with those of control group( P < 0. 05). The membrane potential of mitochondria in Cur group was significantly lower than that in control group( P < 0. 05) in a dose-dependent manner. The expression level of caspase-3 protein in Cur group was significantly higher than that in control group( P <0. 05) in a dose-dependent manner. The expression level of p-AKT protein in Cur group was significantly lower than that in control group( P < 0. 05). Conclusion Cur may induce the apoptosis of U87 cells by means of reducing the mitochondrial membrane potential,initiating the intrinsic mitochondrial apoptotic pathways,activating caspase-3 protein and reducing p-AKT protein.