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目的探讨糖酵解抑制剂3-溴丙酮酸(3-BrPA)诱导乳腺癌细胞SK-BR-3凋亡的作用及其可能相关机制。方法 MTT法检测3-BrPA对乳腺癌细胞SK-BR-3增殖的抑制作用;碘化丙啶(PI)单染流式细胞术检测细胞凋亡;ATP检测试剂盒检测细胞内ATP水平;DHE荧光探针标记法检测细胞内超氧阴离子水平;线粒体膜电位检测试剂盒(JC-1)检测细胞线粒体膜电位。结果MTT结果显示,3-BrPA可抑制SK-BR-3细胞的增殖活性,且呈浓度和时间依赖性。80、160、320μmol·L-1的3-BrPA诱导SKBR-3细胞24 h的凋亡率分别为6.7%、22.3%、79.6%。80、160、320μmol·L-13-BrPA作用于SK-BR-3细胞5 h后,细胞内ATP水平与对照组相比分别为87.7%、60.6%、23.7%。160μmol·L-1的3-BrPA增加SK-BR-3细胞中活性氧的生成,同时使细胞线粒体膜电位降低。结论 3-BrPA可以抑制乳腺癌细胞SK-BR-3的增殖活性,引起线粒体膜电位降低,并且诱导其凋亡,机制可能与抑制肿瘤细胞的糖酵解从而减少ATP的产生并升高细胞内活性氧的水平有关。
Objective To investigate the effect of 3-BrPA on the apoptosis of SK-BR-3 breast cancer cells and its possible mechanism. Methods The inhibitory effect of 3-BrPA on the proliferation of SK-BR-3 breast cancer cells was detected by MTT assay. Apoptosis was detected by flow cytometry with propidium iodide (PI) The level of superoxide anion in cells was detected by fluorescent probe labeling method. The mitochondrial membrane potential was detected by the mitochondrial membrane potential detection kit (JC-1). Results MTT results showed that 3-BrPA could inhibit the proliferation activity of SK-BR-3 cells in a concentration-and time-dependent manner. The apoptosis rates of SKBR-3 cells treated with 80,160,320μmol·L-1 3-BrPA for 24 h were 6.7%, 22.3% and 79.6%, respectively. After treated with 80, 160, 320μmol·L-13-BrPA for 5 h, the level of intracellular ATP was 87.7%, 60.6% and 23.7% compared with the control group, respectively. 3-BrPA at 160 μmol·L-1 increased the production of reactive oxygen species (ROS) in SK-BR-3 cells and decreased the mitochondrial membrane potential. Conclusions 3-BrPA can inhibit the proliferation of SK-BR-3 breast cancer cells and cause a decrease in mitochondrial membrane potential and induce its apoptosis. The mechanism may be that it inhibits the glycolysis of tumor cells to reduce the production of ATP and increase intracellular Reactive oxygen species level.