MEK抑制剂AZD8330对多发性骨髓瘤抗肿瘤作用的初步研究

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.

Effect of AZD8330 on Proliferation and Apoptosis of Multiple Myeloma Cells

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.