氟奋乃静通过自噬诱导U87胶质瘤细胞死亡及其机制探讨

摘要

目的：观察氟奋乃静（FPZ）通过自噬诱导U87胶质瘤细胞死亡的机制。方法体外培养U87胶质瘤细胞并经FPZ处理，采用细胞活力和集落形成方法分析细胞的生存率，采用Western blot检测自噬相关蛋白及PI3K/AKT/mTOR通路蛋白的表达。结果 FPZ处理后，U87胶质瘤细胞活力显著降低（P﹤0.05），菌落数显著降低（P﹤0.05）；微管相关蛋白1轻链3-Ⅱ型（LC3-Ⅱ）显著增加，并呈时间和剂量依赖性；p-AKT水平及其下游p-mTOR水平显著下降，并呈时间依赖性。采用PI3K抑制剂LY294002处理U87胶质瘤细胞后，LC3-Ⅱ生成明显增加。结论 FPZ可通过抑制PI3K/AKT/mTOR通路调节自噬，发挥细胞毒性作用。%Objective To investigate the role of autophagy in fluphenazine (FPZ)-induced cytotoxicity in U87 glio-ma cells. Method U87 glioma cell was cultured in vitro and then treated with FPZ; The survival rate of U87 glioma cells was assessed by cell viability and clonogenic survival assay;The autophagy related protein and PI3K/AKT/mTOR pathway protein were determined by using western blot. Result FPZ treatment inhibited cell viability and long-term clo-nogenic survival of U87 glioma cells (both P<0.05). Additionally, FPZ triggered autophagy, as indicated by the time-and dose-dependent accumulation of microtubule-associated protein 1 light chain 3 (LC3-II), while the level of p-AKT and the downstream p-mTOR was decreased as time elapsed. Treatment with LY294002, a PI3K inhibitor, increased the accu-mulation of LC3-II. Conclusion These results provided the evidence that FPZ-induced cytotoxicity is mediated through autophagic cell death in U87 glioma cells by inhibiting PI3K/AKT/mTOR pathway.