去水卫矛醇对人脑胶质瘤细胞BT325增殖、凋亡及线粒体膜电位的影响

Effects of Dianhydrogalactitol on cell proliferation and apoptosis of human glioma BT325 cells and alternation in mitochondrial membrane potential

目的观察去水卫矛醇(DAG)对人脑胶质瘤BT325细胞增殖、凋亡的影响,探讨其可能的作用机制。方法将BT325细胞分为对照组、不同浓度DAG组,作相应处理后培养,测算细胞存活率、单细胞克隆形成率、细胞凋亡率及线粒体膜电位。结果随DAG浓度的增加,BT325细胞的存活率逐渐下降(P<0.05),不同浓度DAG组随培养时间的延长BT325细胞的存活率逐渐下降(P均<0.05);培养24、48、72 h时,DAG对BT325细胞的IC50分别为108.81、26.61、11.38μg/m L。不同浓度DAG组BT325细胞克隆形成率均为0,与对照组的16.78%±1.50%相比,P均<0.01。5、10、20、40、80μg/m L的DAG组BT325细胞凋亡率分别为11.79%±2.91%、15.42%±3.06%、38.70%±1.12%、60.78%±1.30%、80.35%±3.22%,对照组为4.27%±1.72%;不同浓度DAG组BT325细胞凋亡率与对照组相比,P均<0.01,且呈浓度依赖性(P均<0.01)。5、10、20、40μg/m L的DAG组BT325细胞线粒体膜电位分别为0.174±0.038、0.142±0.037、0.138±0.033、0.119±0.013,对照组为0.193±0.014;20μg/m L的DAG组与对照组相比,P<0.05;40μg/m L的DAG组与对照组相比,P<0.01。结论 DAG抑制BT325细胞增殖,诱导细胞凋亡,作用呈浓度依赖性,其机制可能与降低细胞线粒体膜电位有关。

Objective To investigative the effects of dianhydrogalacitol（DAG）on cell proliferation and apoptosis of human glioma BT325 cells and its potential mechanism.Methods BT325 cells were divided into the control group and DAG-treated groups at various oncentrations.CCK-8 assay and colony formation assay were performed to detect the cell via-bility of BT325 cells.Hoechst33342 was employed to observe nucleus morphological changes and apoptotic rate.The mito-chondrial membrane potential （MMP）was detected by using Rhodamine 123 staining.Results With the increasing DAG concentration,the survival rate of BT325 cells decreased gradually （P〈0.05）.The survival rate of BT325 cells decreased with the prolonged incubation time in DAG-treated groups （all P〈0.05）.The IC50 of BT325 cells treated by DAG were 108.81,26.61,and 11.38 μg/mL at 24,48,and 72 h,respectively.The result of colony formation assay showed that DAG inhibited clones of BT325 cells and the colony formation rate in the control group was 16.78% ±1.50% （all P 〈0.01）.The apoptotic rates of DAG-treated groups treated with 5,10,20,40 and 80 μg/mL DAG were 11.79% ± 2.91%,15.42% ±3.06%,38.70% ±1.12%,60.78% ±1.30%,and 80.35% ±3.22%,respectively,which were in an dose-dependent manner and were significantly increased as compared with that （4.27% ±1.72%）of the control group （all P〈0.01）.The MMP of control group and DAG-treated groups（5,10,20,and 40 μg/mL）were 0.193 ±0.014, 0.174 ±0.038,0.142 ±0.037,0.138 ±0.033,and 0.119 ±0.013,respectively.Significant difference was found in the MMP between the DAG-treated groups （20 and 40 μg/mL）and the control group （P〈0.05 and P〈0.01）.Conclusions DAG inhibits the proliferation and induces apoptosis of BT325 cells.It is suggested that decreased MMP levels might in-volve in the mechanism.