蛋白酶体抑制剂MG132诱导caspase-8依赖的骨肉瘤细胞凋亡机制研究  

Caspase-8 dependent osteosarcoma cell apoptosis induced by proteasome inhibitor MG132

作　　者：严晓波[1] 高翔[1] 冯洁[2] 师钟丽[2] 杨迪生[1] 

机构地区：[1]浙江大学医学院附属第二医院骨科,浙江杭州310009 [2]浙江大学骨科研究所,浙江杭州310009

出　　处：《中国病理生理杂志》2008年第2期231-236,共6页Chinese Journal of Pathophysiology

摘　　要：目的:探讨蛋白酶体抑制剂Z-LLL-CHO(MG132)对人骨肉瘤细胞MG-63的作用以及可能作用机制。方法:将不同浓度的MG132作用于骨肉瘤细胞MG-63,采用显微镜观察形态改变、电镜观察细胞超微结构、MTT检测细胞增殖活力、琼脂糖凝胶电泳检测细胞凋亡、流式细胞仪分析细胞凋亡率以及细胞周期改变、RT-PCR检测基因转录、Western blotting检测蛋白表达水平。结果:MG132能有效诱导人骨肉瘤细胞系MG-63细胞周期在G2-M期的停滞,并引起MG-63的凋亡,出现凋亡的典型形态学改变,同时出现p27kip1蛋白的积累,caspase-8活化蛋白表达增加,Bax∶Bcl-2蛋白含量比例的增高,而对正常的人二倍体成纤维细胞,MG132并未表现诱导其凋亡的活性。结论:MG132引起的人骨肉瘤细胞MG-63的凋亡也许是与caspase-8、p27kip1和Bcl-2蛋白相关的。AIM ： To investigate the effects of proteasome inhibitor Z - LLL - CHO （MG132） on human osteosarcoma cell line MG- 63 and its possibly mechanism. METHODS： After treated with different concentration of MG132, the morphological change, uhrastructral morphology, cell viability, cell apoptosis, gene transcription and protein expression in MG -63 cells were accessed by fluorescence microscope, electron microscope, MTT assay, agrose gel electrophoresis, FCM, RT- PCR and Western blotting. RESULTS： Proteasome inhibitor MG132 was an effective inducer of apoptosis in human osteosarcoma MG - 63 cells. Not only apoptotic changes, but also cell arrest at G2 - M - phase, the accumulation of p27^kip1, the accumulation of activated caspase- 8 and increased ratio of Bax： Bcl -2 were observed. However, to normal human diploid fibroblast cells, MG132 did not show apoptosis -inducing effect. CONCLUSION： Apoptosis induced by MG132 may be caspase - 8, p27^kip1 and bcl - 2 - related.

关 键 词：蛋白酶抑制剂 骨肉瘤 细胞凋亡 蛋白质p27^Kip1 半胱氨酸天冬氨酸蛋白酶8 

分 类 号：R363[医药卫生—病理学]