Hsp90选择性抑制剂17-AAG诱导白血病细胞株凋亡的基因谱研究  

Gene Expression Profile of Apoptosis in Leukemia Cells Induced by Hsp90 Selective inhibitor 17-AAG

作　　者：王娜娜[1] 李之珩[1] 陶燕芳[1] 徐利晓 潘健[1] 胡绍燕[1] 

机构地区：[1]苏州大学附属儿童医院血液科,江苏苏州215006

出　　处：《中国实验血液学杂志》2016年第3期672-680,共9页Journal of Experimental Hematology

基　　金：国家自然科学基金(81170513);国家自然科学基金(81370627);省自然科学基金(BL2013014)

摘　　要：目的:探讨热休克蛋白90(Hsp90)抑制剂17-AAG对急性白血病细胞株HL-60和NB4细胞的凋亡诱导作用及其可能机制。方法:应用CCK-8比色法观察17-AAG对HL-60和NB4细胞的生长抑制作用;用Annexin V/PI标记流式细胞术分析细胞凋亡;Western blot检测凋亡相关蛋白Caspase-3和PARP的表达水平及其活化状态,采用实时定量PCR法检测17-AAG处理后NB4细胞的凋亡相关基因的mRNA变化。结果:17-AAG呈剂量依赖性抑制白血病细胞的生长。Annexin V测定、细胞周期分析和Caspase-3、PARP1的活化均表明17-AAG诱导白血病细胞的凋亡。实时荧光定量PCR分析发现,17-AAG处理后实验组和对照组相比有56个基因显著上调,23个基因显著下调。结论:17-AAG可诱导白血病细胞的凋亡,抑制细胞增殖。17-AAG处理白血病细胞后凋亡相关基因发生明显变化,激活凋亡信号通路可诱导细胞发生凋亡。Objective: To investigate the apoptotic effects of Hsp90 selective inhibitor 17-AAG on human leukemia HL-60 and NB4 cells and analyse its possible mechanism. Methods: CCK-8 assay w as used to quantify the grow th inhibition of cells after exposure to 17-AAG for 24 hours. Flow cytometrve w ith annexin V / propidium iodide staining w as used to detect apoptosis of leukemia cells. Then Western blot w as used to detect the activation of apoptosis related protein caspase-3 and PARP level. Gene expression profile of NB4 cells treated w ith 17-AAG w as analyzed w ith realtime PCR arrays. Results: The inhibition of leukemia cell proliferation displayed a dose-dependent manner. Annexin V assay,cell cycle analysis and activation of PARP demonstrate that 17-AAG induced apoptosis leukemia cells. Real-time PCR array analysis show ed that expression of 56 genes significantly up-regulated and expression of 23 genes w ere significantly dow n-regulated after 17-AAG treatment. Conclusion: The 17-AAG can inhibit the proliferation and induce the apoptosis of leukemia cells. After leukemia cells are treated w ith 17-AAG,the significant changes of apoptosisrelated genes occured,and the cell apoptosis occurs via activating apoptosis related signaling pathw ay.

关 键 词：17-AAG HSP90 细胞凋亡 白血病细胞 

分 类 号：R733.7[医药卫生—肿瘤]