桑葚花色苷提取物对乳腺癌细胞凋亡及线粒体膜电位的影响

Effect of Anthocyanin Extract of Mulberry Fruit on the Apoptosis and the Mitochondrial Membrane Potential of Breast Cancer Cells

目的:观察桑葚花色苷提取物对人乳腺癌细胞株MDA-MB-453、MDA-MB-231和MCF-7细胞凋亡及线粒体膜电位的影响。方法:利用超声辅助乙醇萃取法提取桑葚花色苷,pH示差法测定提取物花色苷总含量,以50、100和150 mg/mL桑葚花色苷提取物作用三种乳腺癌细胞MDA-MB-231、MDA-MB-453和MCF-7 24 h,采用Annexin V/PI双染流式细胞分析法检测细胞凋亡水平变化,JC-1探针染色激光共聚焦扫描显微镜观察MDA-MB-453细胞线粒体膜电位水平变化。结果:凋亡分析结果表明,桑葚花色苷提取物作用后三种乳腺癌细胞凋亡率均升高,显示出促凋亡效应,且具有剂量-效应关系,100和150 mg/mL组凋亡率显著升高(P<0.05)。激光共聚焦扫描显微镜检测结果显示,桑葚花色苷提取物作用24 h,可使MDA-MB-453细胞线粒体膜电位显著下降,表现为红色/绿色荧光的比值显著降低(P<0.05)。结论:桑葚花色苷提取物可显著降低乳腺癌细胞线粒体膜电位,并促发细胞凋亡。

Objective：To study the effect of anthocyanin extract of mulberry fruit on the apoptosis and the mitochondrial membrane potential of breast cancer cells MDA-MB-453,MDA-MB-231 and MCF-7.Methods：Preparation of anthocyanin-rich extract from mulberry fruits was carried out by ultrasonic extraction with acidified-ethanol.The breast cancer cells MDA-MB-453,MDA-MB-231 and MCF-7 were reated with 50,100 or 150 mg/mL of the anthocyanin-rich extract for 24h;the cells apoptosis were analyzed by Annexin V/PI dyeing and flow cytometric assay.To characterize the upstream factors involved in the intrinsic apoptosis pathway,the mitochondrial permeability of MDA-MB-453 cells were measured by JC-1 staining and the laser confocal scanning microscopy.Results：The apoptosis analysis results indicated that the anthocyanin-rich mulberry fruit extract treated for 24 h,the apoptosis rate of these three breast cells in 100 and 150 mg/mL treated groups markedly increased（P0.05）,exhibiting a dose-effect relationship.The observation by laser confocal scanning microscopy suggested that the anthocyanin-rich extract significantly decreased the mitochondrial membrane potential of breast cancer MDA-MB-453 cell;the ratio of JC-1 red/green fluorescence was significantly decreased（P0.05）.Conclusion：The anthocyanin-rich extract significantly decreased the mitochondrial membrane potential of breast cancer cells and induced apoptosis.