VX-680体外诱导异基因干细胞移植后复发AML细胞凋亡的初步研究  

Apoptosis induced by VX-680 in leukemia cells from AML patients relapsed after allogeneic stem cell transplantation in vitro

作　　者：许多荣[1] 赖淑萍[1] 邹外一[1] 黄珊[1] 李娟[1] 

机构地区：[1]中山大学附属第一医院血液科,广东广州510080

出　　处：《中华肿瘤防治杂志》2011年第19期1501-1505,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30670997);广东省科技计划项目(2011B031800120)

摘　　要：目的:探讨Aurora激酶抑制剂VX-680在体外对异基因造血干细胞移植(Allo-HSCT)后复发的急性髓系白血病(AML)患者细胞增殖和凋亡的影响。方法:以正常人CD34+细胞为对照,体外用5nmol/L浓度VX-680分别处理3例Allo-HSCT后复发的AML患者白血病细胞,48h后用四甲基偶氮唑蓝(MTT)法观察细胞增殖活力,Hoechest33342方法观察细胞凋亡形态,流式细胞术检测细胞凋亡百分率,蛋白质印迹法检测Aurora-A磷酸化蛋白及Caspase-3凋亡蛋白的表达。结果:MTT法观察发现,体外VX-680均能抑制3例复发患者白血病细胞的增殖,增殖抑制率分别为(77.5±3.1)%、(76.8±4.7)%和(81.1±4.2)%,均明显高于正常对照的(11.2±1.6)%,P值均<0.001;Hoe-chest33342方法观察发现,3例患者白血病细胞在VX-680作用下出现了明显的细胞凋亡形态,流式细胞术检测到凋亡百分率分别为(88.8±4.6)%、(98.7±0.9)%和(99.3±0.4)%,也均高于正常对照的(13.6±3.5)%,P值均<0.001;蛋白质印迹法检测显示,VX-680处理后3例患者白血病细胞中Aurora-A磷酸化蛋白表达显著下降,而Caspase-3凋亡蛋白表达明显升高。结论:VX-680体外可通过诱导Allo-HSCT后AML复发患者的白血病细胞凋亡来抑制其增殖。OBJECTIVE： To investigate the effect of Aurora kinase inhibitor,VX-680,on the proliferation and apoptosis in leu kemia cells from AML patients relapsed after allogeneic stern cell transplantation（Allo HSCT） in vitro. METHODS： Using CD34＋ cells from a health people as control,CD34＋ leukemia cells from 3 cases of AML patients relapsed after Allo-HSCT were incubated respectively with VX-680 at 5 nmol/L for 48 h in vitro. The proliferation capacity was measured by MTT assay,the morphology of cell apoptosis was observed with Hoechst33342 staining by fluorescence microscopy, the percentage of apoptosis was calculated by flow cytometry, and the expression of the phosphorylated protein AurormA and the apoptosis protein Caspase-3 was determined by Western blot. RESULTS： The proliferation of leukemia cells from the 3 relapsed cases was inhibited by VX-680 in vitro ,and the incidence of inhibition were （77.5±3. 1）%, （76.8±4. 7）% and （81.1± 4. 2）% respectively, which were significantly higher than （11.2!1.6） % in control group（P〈0. 001）. The morphology of leukemia cells from the 3 cases apparently showed apoptosis after treated with VX-680, and the percentage of apoptosis could be up to （88.8±4.6）%,（98.7±0.9）% and （99.3±0.4）% respective- ly,which were higher than （ 13.6± 3.5） % in control group（P〈 0. 001）. It was also confirmed that the expression of the phospho rylated protein Aurora-A were down-regulated and the apoptosis protein Caspase 3 were up-regulated significantly in leukemia cells from the 3 cases after treated with VX-680 by Western blot assay. CONCLUSION： VX-680 can inhibit the proliferation of leukemia cells from AML patients relapsed after Allo-HSCT in vitro by inducing cell apoptosis.

关 键 词：白血病 髓样 急性 VX-680 造血干细胞移植 复发 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]