维生素E琥珀酸酯诱导MDA-MB-435乳腺癌细胞的凋亡

Vitamin E succinate induced apoptosis of MDA-MB-435 human breast cancer cells

目的:检测维生素E琥珀酸酯(vitaminEsuccinate,VES)对MDAMB435乳腺癌细胞增殖抑制和凋亡诱导作用并分析凋亡诱导分子Fas表达的变化。方法:以VES刺激人乳腺癌细胞MDAMB435(雌激素受体阴性)12、24和48h,VES浓度为5、10和20μg/mL,以MTT法测定对细胞增殖的抑制作用,以流式细胞仪分析细胞周期和细胞表面Fas表达,TUNEL法检测肿瘤细胞凋亡指数变化,Westernblot法检测VES作用后Fas蛋白水平变化。结果:VES对MDAMB435乳腺癌细胞增殖具有显著的抑制作用并表现为时间和剂量依赖关系。流式细胞仪分析细胞凋亡,MDAMB435细胞的自然凋亡率为1.5%,5、10和20μg/mLVES分别作用48h后凋亡率升高至13.1%、20.2%和46.5%。TUNEL法检测VES作用后乳腺癌细胞凋亡指数由1.1±0.4升高至10.8±1.0、30.4±2.7和51.4±4.7,细胞Fas蛋白水平分别升高1.3、1.9和4.3倍,细胞表面Fas平均荧光强度由46.29升高至59.66、84.54和103.41。结论:VES对MDAMB435乳腺癌细胞具有显著的增殖抑制作用,并诱导细胞的凋亡,其机制与细胞表面Fas表达上调有关。

Objective：To investigate the growth inhibition and apoptosis induction effect of vitamin E succinate （VES） on MDA MB-435 human breast cancer cells and to analyze the changes of Fas expression in this process. Methods： MDA-MB-435 human breast cancer cells were treated with VES at concentrations of 5μg/mL, 10μg/mL, and 20μg/mL for 12, 24, and 48 h, respectively. The inhibitory effect of VES on cell proliferation was measured with MTT assay. Cell cycle and cell surface Fas ex pression were analyzed by flow cytometry. Apoptotie index was measured by TUNEL method. Fas protein level was detected by Western blotting assay. Results：YES had significant inhibitory effect on the growth of MDA-MB-435 human breast cancer cells. The inhibitory effect was in a time- and dose-dependent manner. Flow cytometry analysis showed that the apoptotic rate rose from 1.5% to 13.1% ,20.2% ,and 46.5% after treatment with VES for 48 h at the concentrations of 5μg/mL, 10μg/mL,and 20μg/mL,respectively. Both apoptotic index and Fas expression of MDA-MB-435 ceils increased after the treatment with VES. Apoptotie index rose from 1.1±0.4 to 10.8±1.0,30.4±2.7,and 51.4±4.7 after VES treatment. Fas protein level increased increased 1.3, 1.9 and 4.3 folds, respectively. The mean fluorescence intensity of Fas on cell surface rose from 46.2 to 59.66, 84.54, and 103.41, Cunelusion： VES had significant growth inhibition and apoptosis induction effect on MDA-MB 435 estrogen receptornegative breast cancer cells. The mechanism involved up-regulation of Fas on the surface of cancer cells.