MEK抑制剂AZD8330对多发性骨髓瘤抗肿瘤作用的初步研究  

Effect of AZD8330 on Proliferation and Apoptosis of Multiple Myeloma Cells

作　　者：姚瑶[1] 边月平 夏丹丹[1] 潘彬[1] 牛铭山[1] 赵恺[1] 曾令宇[1] 徐开林[1] 

机构地区：[1]徐州医学院附属医院血液科,江苏徐州221006

出　　处：《中国实验血液学杂志》2014年第5期1311-1315,共5页Journal of Experimental Hematology

基　　金：国家自然科学青年基金(81302034)

摘　　要：本研究探讨MEK抑制剂AZD8330对多发性骨髓瘤细胞IM9及NCI-H929细胞增殖、凋亡的影响及其可能的作用机制。使用不同浓度AZD8330处理NCI-H929及IM9细胞48 h,用CCK-8检测细胞活力,并计算出48 h的IC50;分别用5、10及100 nmol/L的AZD8330处理上述两种细胞,然后用PI标记流式细胞术分析细胞周期的变化;Western blot方法检测细胞周期相关蛋白cyclin D及cyclin E;分别用10、100、1000及2000 nmol/L的AZD8330处理骨髓瘤细胞,AnnexinV/7-AAD双标记法分析细胞凋亡,并应用Western blot方法检测凋亡相关蛋白caspase-3。结果表明,AZD8330可显著抑制NCI-H929及IM9的细胞活力,且抑制效应呈一定的时间和剂量依赖性,IM9细胞48 h的半数抑制浓度(IC50)为19.88±2.7 nmol/L,NCI-H929细胞48 h的半数抑制浓度(IC50)为29.3±2.03 nmol/L。细胞周期结果显示,两种骨髓瘤细胞均发生G1期阻滞;AZD8330处理后cyclin D水平显著增高,但是cyclin E水平降低,促使细胞发生G1期阻滞。AnnexinV/7-AAD结果显示,随着AZD8330浓度增加,细胞凋亡数量也相应增多,并且活化的caspase-3蛋白水平增高。结论:MEK抑制剂AZD8330可有效抑制骨髓瘤细胞NCI-H929和IM9的增殖,使细胞周期阻滞于G1期,并促进细胞发生凋亡。This study was aimed to investigate the effect of MEK inhibitor AZD8330 on proliferation and apoptosis of multiple myeloma IM9 and NCI-H929 cell lines and its possible mechanism.These two cell line cells were exposed to different concentrations of AZD8330 for 48 h.The CCK-8 assay was used to detect cell,viability and the IC50 value at 48 h.These above-mentioned IM9 and NCI-H929 cells were treated with 5,10 and 100 nmoL/L of AZD8330,then the change of cell cycle was analysed by flow cytometry with PI staining.The Wester blot was used to detect the expression levels of cyclin D and cyclin E,and multiple myeloma cells were treated with 10,100,1000 and 2000 nmol/L of AZD8330,the AnnexinV/7-AAD double staining was used to analyse cell apoptosis and the Western blot was used to detect the expression level of caspase-3.The results showed that AZD8330 could significantly inhibit the cell viability of IM9 and NCI-H929 cell lines in a time-and dose-dependent manner,the IC50 value （48 h） of IM9 and NCI-H929 were 19.88 ± 2.7 nmol/L and 29.3 ± 2.03 nmol/L respectively,these two cell lines were arrested on G1 phase of cell cycle,the apoptosis cells increased along with enhancement of AZD8330 concentration,and the expression level of cleaved caspase-3 protein was up-regulated.It is concluded that AZD8330 can efficiently inhibit the proliferation of NCI-H929 and IM9 cell lines,and induce apoptosis,suggesting that the AZD8330 may be a potential chemotherapeutic candidate for multiple myeloma therapy.

关 键 词：MEK抑制剂 AZD8330 多发性骨髓瘤 细胞凋亡 细胞周期 

分 类 号：R733.3[医药卫生—肿瘤]