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

摘要

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／mL。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／mL 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／mL 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／mL 的 DAG 组与对照组相比，P ＜0．05；40μg／mL 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.