双氢青蒿素对恶性胶质母细胞瘤细胞增殖与凋亡的影响

Dihydroartemisinin Inhibits Proliferation and Induces Apoptosis of U87 Cells by Increasing the Expression of Caspase-3

目的探讨双氢青蒿素(DHA)对人恶性胶质母细胞瘤(U87)细胞增殖和凋亡的影响。方法将10,25,50,100,125μmol.L-1DHA分别作用于体外常规培养的U87细胞24,48和72 h,CCK-8细胞增殖试剂盒实验检测细胞增殖情况并计算DHA不同作用时间半数抑制浓度(IC50);97.04μmol.L-1DHA作用U87细胞24 h后倒置显微镜观察细胞形态变化,流式细胞分析细胞周期变化,Hoechst 33258染色观察细胞凋亡,检测Caspase-3蛋白含量变化,双氢-乙酰乙酸二氯荧光黄(DCFH-DA)荧光探针检测活性氧(ROS)变化。结果 DHA浓度、作用时间不同,U87细胞存活率不同,差异有统计学意义(P<0.05),DHA对U87细胞的抑制作用呈浓度和时间依赖性。DHA作用24 h时IC50为97.04μmol.L-1,48 h为59.76μmol.L-1,72 h为33.20μmol.L-1;97.04μmol.L-1DHA作用24 h后S期细胞减少,G0～G1期细胞增多,凋亡细胞增多,细胞内ROS增多。Caspase-3表达量较对照组高,差异有统计学意义(P<0.05)。结论 DHA对U87增殖有抑制作用,可诱导细胞凋亡。其机制可能与DHA改变细胞周期,激活Caspase-3通路以及增加细胞内ROS有关。

Objective To investigate the effect of dihydroartemisinin （DHA） on apoptosis and proliferation of U87 ceils. Methods U87 cells were cultured with DHA （ 10,25,50,100, and 125 μmol·L-1 ） for 24,48 and 72 h. Cell counting （CCK-8） assay was employed to evaluate the survival of DHA-treated U87 cells. The induction of apoptosis was detected by Hoechst 33258. Effect of DHA on cell cycle of U87 cells was detected by flow cytometry. Caspase-3 activities were measured with Caspase-3 activity assay kit and reactive oxygen species （ROS） measured with reactive oxygen species assay kit. Results The results indicated that DHA induced apoptotie eel1 death in a dose- and time-dependent manner,which was accompanied by the activation of Caspase-3 and increased ROS. The IC50 was 97.04μmol·L-1 ,59.76 μmol·L-1 and 33.20 μmol·L-1 at 24, 48, and 72 h, respectively. The apoptosis cells and Go to G1 phase cells increased after treated with 97.04 μmol·L-1 DHA, while the S phase cells decreased. Meanwhile, the expression of Caspase-3 was higher in the DHA treatment group compared with control group （P〈0.05）. Conclusion Dihydroartemisinin exerts eytotoxic effect on U87 cells by altering the cell cycle, activating Caspase-3 activities and increasing the ROS.