U266细胞在组蛋白去乙酰化酶抑制剂MS-275诱导下凋亡观察
Effects of histone deacetylase inhibitor MS-275 in vitro on apoptosis of U266 cell line
马健 1赵名 2于晓妉 2王志红1
作者信息
- 1. 解放军总医院第一附属医院血液科,北京 100037
- 2. 军事医学科学院基础医学研究所,北京 100850
- 折叠
摘要
目的:研究组蛋白去乙酰化酶抑制剂MS-275对人多发件骨髓瘤U266细胞的增殖和凋亡的影响.方法:台盼蓝拒染法观察MS-275对细胞活力的影响;瑞氏-姬姆萨染色观察药物对细胞形态学变化;流式细胞仪分析细胞周期;Western Blot检测凋亡信号通路中Caspase-3,Caspase-8,Caspase-9活化及蛋白聚ADP核糖聚合酶[poly(ADP-ribose)polymerase,PARP]裂解情况.结果:MS-275呈时间和剂量依赖性抑制U266细胞增殖,阻断细胞周期于G0/G1期.MS-275作用48 h的IC50为1.39 μmol/L;2 μmol/L MS-275作用细胞24 h后,G0/G1期64.57%;36 h占82.20%.瑞氏-姬姆萨染色显示细胞发生明显变化.Western Blot检测表明MS-275作用U266细胞后,Caspase-3,Caspase-8,Caspase-9被裂解活化,其底物蛋白聚ADP核糖聚合酶发生剪切,细胞发生凋亡.结论:MS-275可抑制细胞增殖,并使细胞阻滞在G0/G1期,通过激活细胞内外2条凋亡级联信号通路诱导U266细胞凋亡.
Abstract
AIM: To study the effect of MS-275,an inhibitor of histone deacetylase,on the growth and apoptosis of myeloma cell line U266. METHODS: U266 cell was cultured in RPMI-1640 in the presence of MS-275 and the cell viability was evaluated by Trypan blue exclusion assay and cell count. The cell morphological change was observed with Wright-Giemsa staining. The cell cycle was analyzed by flow cytometry and the proteins of poly ( ADP-ribose ) polymerase ( PARP ),Caspase-3,Caspase-8 and Caspase-9 were detected by Western Blot. RESULTS: MS-275 inhibited the growth of U266 cells in a dose-and time-dependent manner. The cell cycle was arrested at G0/G1 phase. After exposure to 1. 39 μmol/L MS-275 for 48 h,the cell viability decreased to 50%. After 2 μmol/L MS-275 treatment for 24 h and 36 h,the cell ratios of G0/G1 phase increased respectively to 64. 57% and 87.20%. The morphological visible changes of U266 were confirmed by Wright-Giemsa staining. The proteins were abstracted from the treated cells,and Caspase-3,Caspase-8 and Caspase-9 activation and cleavage of PARP were examined by Western blot using a specific antibody. The cleaved Caspase-3,Caspase-8 and Caspase-9 and cleaved PARP were recognized respectively. CONCLUSION: MS-275 inhibits the proliferation and arrests the cell cycle at G0/G1 phase. MS-275 induces apoptosis of U266 cells through both extracellular and intracellular signaling cascade pathways.
关键词
组蛋白去乙酰化酶抑制剂/多发性骨髓瘤/细胞系U266/MS-275/凋亡Key words
histone deacetylase inhibitors/multiple myeloma/cell line U266/MS-275/apoptosis引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据