姜黄素诱导自噬抑制脑胶质瘤U87细胞增殖的研究

目的研究姜黄素对脑胶质瘤U87细胞增殖的抑制作用及机制。方法采用10、20、40、60、80μmol/L姜黄素分别处理U87细胞72h后,MTT法检测姜黄素对U87细胞增殖的影响。流式细胞术检测40μmol/L姜黄素处理前后U87细胞周期和凋亡变化。提取40μmol/L姜黄素处理前后不同时间点U87细胞总蛋白,Westernblot检测自噬相关蛋白LC3的表达。结果姜黄素呈剂量依赖性抑制U87细胞增殖(P0.05),半数抑制浓度(IC50)为42.44μmol/L。姜黄素诱导G2-M期细胞周期阻滞,实验组G2-M期细胞比例为(47.71±2.67)%,对照组为(17.96±0.88)%(P0.01);实验组细胞凋亡率为(3.93±0.82)%,对照组为(1.80±0.65)%,两组差异无统计学意义(P0.05)。姜黄素给药12、24、48h,LC3-Ⅱ/LC3-Ⅰ比值分别为(0.89±0.004)、(0.93±0.01)、(1.09±0.02),高于对照组(0.76±0.04)(P0.05);3-MA能减弱姜黄素对U87细胞增殖的抑制作用。结论姜黄素通过诱导自噬抑制脑胶质瘤U87细胞的增殖。

Inhibitory effect of curcumin on the proliferation of malignant glioma cells U87 by inducing autophagy

Objective To investigate the inhibitory effect and potential mechanism of curcumin on the proliferation of U87 cells. Methods After being treated with 10,20,40,60,80 μmol/L curcumin respectively for 72 h,the effect of curcumin on the proliferation of U87 cells was examined by MTT assay. Cell cycle and apoptosis of U87 cells were evaluated by flow cytometry （FCM） before and after being treated with 40 μmol/L curcumin. After extracting total protein from U87 cell at different time points,the expression of LC3 was examined by Western blotting. Results Curcumin displayed a growth inhibitory effect on U87 cells in a dose-dependent manner （P 0.05）. The IC50 of curcumin was 42.44 μmol/L. The cell cycles analyses indicated that cells were arrested at G2-M phases,and cell percentage in G2-M stage was （47.71 ± 2.67） % in curcumin group and （17.96 ± 0.88）% in blank control （P 0.01）. Apoptosis rate was （3.93±0.82） % in curcumin group,（1.80 ± 0.65）% in blank control,and no significant difference was found between the two groups （P 0.05）. After being treated with curcumin （40 μmol/L） for 12,24,48 h respectively,the ratio values of LC3-Ⅱ/LC3-Ⅰwere 0.89 ± 0.004,0.93 ± 0.01,1.09 ± 0.02 respectively,higher than control group （0.76 ± 0.04） （P 0.05）. 3-MA decreased the inhibitory effect of curcumin on U87 cells. Conclusions Curcumin suppresses the proliferation of malignant glioma cells U87 through induction of autophagy.