肿瘤坏死因子相关凋亡诱导配体诱导胶质瘤细胞凋亡机制研究  

Tumor necrosis factor-related apoptosis-inducing ligand induces glioma U87 cell apoptosis through the mitochondrial apoptotic pathway

作　　者：仲飞[1] 邵春奎[2] 林金香[1] 杨静[3] 吴祥元[1] 林曲[1] 董敏[1] 温景芸[1] 马小琨[1] 魏丽[1] 

机构地区：[1]中山大学附属第三医院肿瘤内科,广州510630 [2]中山大学附属第三医院病理科 [3]中山大学附属第三医院特诊医学中心

出　　处：《中华神经医学杂志》2008年第9期878-880,885,共4页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（30672409）;广东省自然科学基金（05001748）

摘　　要：目的 探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）诱导胶质瘤细胞凋亡的机制。方法以人重组可溶性TRAIL蛋白（rsTRAIL）处理人胶质瘤细胞U87：Annexin V—FITC／PI双染色流式细胞术检测细胞凋亡；DiOC6荧光染色流式细胞术检测细胞线粒体跨膜电位（△ψm）：比色法测定细胞caspase-3、8、9活性的变化；ELISA法检测胞浆cytC浓度；观察caspase-8阻断剂（Z—IETD—fmk）对U87细胞凋亡、△ψm和caspase-3、8、9活性变化的影响。结果rsTRAIL以时间依赖方式诱导U87细胞凋亡。同时导致U87细胞△ψm进行性下降，caspase-3、8、9活性及胞浆内cytC浓度升高：caspase-8阻断剂可明显减弱rSTRAIL对U87细胞的上述生物学效应。结论TRAIL通过激活caspase-8间接启动线粒体凋亡途径诱导恶性胶质瘤细胞U87凋亡。Objective To study the mechanism of tumor necrosis factor-related apoptosis-inducing ligand （TRAIL）-induced apoptosis of glioma U87 cells. Methods Human glioma U87 cells were treated with human recombinant soluble TRAIL （rsTRAIL）, and the cell apoptosis was detected with flow cytometry with AnnexinV-FITC/PI double staining. Flow cytometry with DiOC6 staining was used to assess the changes in mitochondrial transmembrane potential （△ψm）. The relative activity of caspase-3, -8 and -9 was measured by colorimetric assay, and the concentration of cytoplasmic cytochrome c （cyt C） determined using enzyme-linked immunosorbent assay. The effects ofcaspase-8 inhibitor （Z-IETD-fmk） on rsTRAIL-induced apoptosis, △ψm, caspase-3, -8 and -9 activities and cyt C concentration were observed. Results RsTRAIL time-dependently induced apoptosis and progressive collapse of △ψm in glioma U87 cells, resulting also in caspase-3, -8 and -9 activation and elevated cyt C concentration. Caspase-8 inhibitor partially antagonized these biological effects induced by rsTRAIL in U87 cells. Conclusion TRAIL initiates a cascade ofmitochondrial events by activating caspase-8 and induces apoptosis ofglioma U87 cells.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 神经胶质瘤 细胞凋亡 线粒体 

分 类 号：R739.4[医药卫生—肿瘤]