中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2008年
9期
878-880,885
,共4页
仲飞%邵春奎%林金香%杨静%吴祥元%林曲%董敏%温景芸%马小琨%魏丽
仲飛%邵春奎%林金香%楊靜%吳祥元%林麯%董敏%溫景蕓%馬小琨%魏麗
중비%소춘규%림금향%양정%오상원%림곡%동민%온경예%마소곤%위려
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
腫瘤壞死因子相關凋亡誘導配體%神經膠質瘤%細胞凋亡%線粒體
종류배사인자상관조망유도배체%신경효질류%세포조망%선립체
Tumor necrosis factor-related apoptosis-inducing ligand%Glioma%Apoptosis%Mitochondria
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;AnnexinV-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cyt C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 TRAU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.
目的 探討腫瘤壞死因子相關凋亡誘導配體(TRAIL)誘導膠質瘤細胞凋亡的機製.方法 以人重組可溶性TRAIL蛋白(rsTRAIL)處理人膠質瘤細胞U87;AnnexinV-FITC/PI雙染色流式細胞術檢測細胞凋亡;DiOC6熒光染色流式細胞術檢測細胞線粒體跨膜電位(△ψm);比色法測定細胞caspase-3、8、9活性的變化;ELISA法檢測胞漿cytC濃度;觀察caspase-8阻斷劑(Z-IETD-fmk)對U87細胞凋亡、△ψm和caspase-3、8、9活性變化的影響. 結果 rsTRAIL以時間依賴方式誘導U87細胞凋亡.同時導緻U87細胞△ψm進行性下降,caspase-3、8、9活性及胞漿內cyt C濃度升高:caspase-8阻斷劑可明顯減弱rsTRAIL對U87細胞的上述生物學效應. 結論 TRAU通過激活caspase-8間接啟動線粒體凋亡途徑誘導噁性膠質瘤細胞U87凋亡.
목적 탐토종류배사인자상관조망유도배체(TRAIL)유도효질류세포조망적궤제.방법 이인중조가용성TRAIL단백(rsTRAIL)처리인효질류세포U87;AnnexinV-FITC/PI쌍염색류식세포술검측세포조망;DiOC6형광염색류식세포술검측세포선립체과막전위(△ψm);비색법측정세포caspase-3、8、9활성적변화;ELISA법검측포장cytC농도;관찰caspase-8조단제(Z-IETD-fmk)대U87세포조망、△ψm화caspase-3、8、9활성변화적영향. 결과 rsTRAIL이시간의뢰방식유도U87세포조망.동시도치U87세포△ψm진행성하강,caspase-3、8、9활성급포장내cyt C농도승고:caspase-8조단제가명현감약rsTRAIL대U87세포적상술생물학효응. 결론 TRAU통과격활caspase-8간접계동선립체조망도경유도악성효질류세포U87조망.
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 detecmd 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 measmed by colorimetric assay,and the concentration of cytoplasmic cytochrome C(cyt C) determined using enzyme-linked immunosorbem 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 tinle-dependently induced apoptosis and progressive collapse of △ψm in glioma U87 cells,resulting also in caspase-3,-8 and-9activation and elevated cytC concentration.Caspase-8 inhibitor partially antagonized these biological effects induced by rsTRAIL in U87 cells.Conclusion TRAIL initiates a cascade of mitochondrial events by activating caspase-8 and induces apoptosis of glioma U87 cells.
