双氢青蒿素促进胶质瘤GL261细胞的凋亡

Dihydroartemisinin induces apoptosis in GL261 glioma cells

目的研究双氢青蒿素对胶质瘤GL261细胞凋亡的影响与作用机制。方法 0、30、60、90μmol/L双氢青蒿素处理GL261细胞,倒置显微镜下观察细胞形态改变,CCK-8法检测处理后GL261细胞活力的改变,Annexin V PI法检测双氢青蒿素对GL261细胞凋亡的影响,免疫荧光显微镜观察双氢青蒿素处理后激活型Caspase-3的表达改变与亚细胞定位,Western blot检测激活型Caspase-3与抗凋亡蛋白Bcl-xL的表达变化。结果双氢青蒿素能有效诱导胶质瘤GL261细胞凋亡,抑制细胞活性,具有浓度依赖性(P<0.05);30μmol/L双氢青蒿素作用12 h即可诱导Caspase-3激活并与细胞核共定位;双氢青蒿素能够下调Bcl-xL、上调激活型Caspase-3的表达水平,且具有浓度依赖性(P<0.05)。结论双氢青蒿素通过抑制抗凋亡蛋白Bcl-xL、诱导Caspase-3激活途径介导胶质瘤GL261细胞凋亡。

Objective To investigate the role and mechanism of dihydroartemisinin in inducing apoptosis in glioma GL261 cells. Methods GL261 cells were treated with different concentrations of dihydro- artemisinin （0, 30, 60 and 90 μmol/L）. The morphological changes of GL261 were observed by an inverted microscope, and the viability of GL261 cells was measured by CCK-8 assay. Annexin V PI assay was used to study the percentage of apoptotic GL261 cells. Laser scanning confocal microscopy was applied to examine the expression and subcellular localization of activated caspase-3 in the GL261 cells, and Western blotting was carried out to detect the expression of activated caspase-3 and Bcl-xL. Results Dihydroartemisinin reduced the viability and induced apoptosis of GL261 ceils in a dose-dependent manner （ P 〈 0.05 ）. Treatment with 30 μmol/L dihydroartemisinin for 12 h could induce the activation of caspase-3 and co-localization with the nucleus in GL261 cells. Dihydroartemisinin could up-regulate the level of activated caspase-3 and down-regulate the level of Bcl-xL in GL261 cells in a dose-dependent manner （P 〈0. 05）. Conclusion Dihydroartemisinin induces apoptosis in GL261 ceils. Anti-apoptotic protein Bci-xL and the executor caspase-5 are involved in this process.