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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。
AIM: To investigate the effects of Aurora kinase inhibitor VX-680 on proliferation and apoptosis of acute myeloid leukemia (AML) patients relapsed after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) in vitro. Methods: The normal human CD34 + cells were used as control. The leukemic cells of AML patients relapsed after Allo-HSCT were treated with 5nmol / L VX-680 in vitro. After 48h, the cells were treated with MTT assay Cell viability was observed by flow cytometry. The morphology of apoptotic cells was observed by Hoechest 33342 method. The percentage of apoptotic cells was detected by flow cytometry. The expressions of Aurora-A phosphorylation protein and Caspase-3 apoptotic protein were detected by Western blotting. Results: MTT assay showed that VX-680 inhibited the proliferation of leukemia cells in 3 patients with recurrence in vitro. The inhibitory rates of proliferation were (77.5 ± 3.1)%, (76.8 ± 4.7)% and (81.1 ± 4.2)%, respectively Significantly higher than the control (11.2 ± 1.6)%, P values were <0.001; Hoe-chest33342 method was observed in 3 patients with leukemia cells under the action of VX-680 obvious apoptotic morphology, flow cytometry The percentages of apoptotic cells were (88.8 ± 4.6)%, (98.7 ± 0.9)% and (99.3 ± 0.4)%, respectively, also higher than that of the normal controls (13.6 ± 3.5% The results showed that the expression of Aurora-A phosphorylation protein in leukemia cells of VX-680 group was significantly decreased, while the expression of Caspase-3 protein was significantly increased. CONCLUSION: VX-680 inhibits the proliferation of leukemic cells in vitro by inducing AML relapse after Allo-HSCT in vitro.