2-甲氧基雌二醇上调Bax/BCL-2比例诱导淋巴瘤Raji细胞凋亡的研究

Up-Regulation of Bax/BCL-2 Ratio by 2-Methoxyestradiol Induces Apoptosis in Lymphoma Raji Cells

目的:研究2-甲氧基雌二醇(2-ME2)对淋巴瘤Raji细胞的作用及其作用机制。方法:用不同浓度2-ME2作用于淋巴瘤Raji细胞,CCK8法检测2-ME2对Raji细胞增殖的影响;流式细胞仪FITC/PI双标法检测细胞早期的凋亡;蛋白印迹法检测2-ME2对Raji细胞BCL-2、Bax、Caspase-3、C-myc蛋白表达的影响。结果:2-ME2对Raji细胞的增殖具有明显抑制作用,其抑制率随着药物浓度的增高而增加,随着药物作用时间的延长而显著增高(r=0.9215)。流式细胞仪FITC/PI双染色法结果显示,2.5μmol/L 2-ME2处理24 h组的细胞凋亡率为(33.79±1.63)%,而48 h组的凋亡率高达(51.90±2.72)%,对照组Raji细胞为(7.08±0.36)%。2.5μmol/L 2-ME2处理Raji细胞12 h后,蛋白印迹检测结果显示,Bax蛋白表达上调,BCL-2蛋白下调,而Caspase-3蛋白表达亦上调,C-myc蛋白表达下调,之间均具有时效性关系。结论:2-ME2对淋巴瘤Raji细胞具有明显的抑制作用,且呈剂量和时间依赖性。其对淋巴瘤Raji细胞的作用机制可能与上调Bax/BCL-2比例,激活Caspase-3诱导癌细胞凋亡有关,下调C-myc蛋白表达也参与了凋亡过程。

Objective:To investigate the effect of 2-methoxyestradiol(2-ME2)to lymphoma Raj 1 cells and its mechanism.Methods:Different concentrations of 2-ME2 were used to treat lymphoma Raji cells.CCK8 method was used to detect the effect of 2-ME2 to proliferation of Raji cells.Flow cytometry FITC/PI double labeling method was used to detect early apoptosis of the cells.Western blotting was used to detect the effect of 2-ME2 to the expression of BCL-2,Bax,Caspase-3 and C-myc proteins in Raji cells.Results:2-ME2 significantly inhibited the proliferation of Raji cells.The inhibition rate increased with the increasing of drug concentration,and increased significantly with the prolongation of drug treatment time(r=0.9215).Flow cytometry FITC/PI double staining showed that the apoptotic rate of 2.5μmol/L2-ME2 treatment group was(33.79±1.63)%,while the apoptosis rate of the 48 h group was(51.90±2.72)%,and that of the control group was(7.08±0.36)%.After treated with 2.5μmol/L 2-ME2 for 12 h,the expression of Bax protein was up-regulated,BCL-2 protein was down-regulated,caspase-3 protein expression was up-regulated,and C-myc protein expression was down-regulated,all of them showed a time-dependent relationship.Conclusion:2-ME2 shows obvious inhibitory effect on lymphoma Raji cells in a dose-and time-dependent manner.Its mechanism of treatment on lymphoma Raji cells may be related to up-regulation of Bax/BCL-2 ratio and activation of Caspase-3 to induce apoptosis in cancer cells.Down-regulation of C-myc protein expression also participates in the apoptotic process.