摘要
目的:探讨三氧化二砷联合伊曲康唑对急性髓系白血病KG1a细胞株的增殖与凋亡的影响及其可能的机制。方法:用瑞氏姬姆沙染色法观察细胞形态;CCK-8检测细胞抑制率;甲基纤维素集落形成实验检测细胞集落形成能力;流式细胞术检测细胞凋亡率及细胞周期阻滞变化;Western blot检测细胞BCL-2、caspase-3、BAX、SMO、Gli1、Gli2蛋白表达。结果:三氧化二砷和伊曲康唑单用对KG1a细胞均具有增殖抑制作用,且呈时间剂量依赖性。联合用药与单药对比,前者细胞存活率、集落形成数均明显减少(P<0.05),而细胞凋亡率增高。联合用药能够增加细胞的Caspase-3和BAX两种蛋白的表达、下调BCL-2、SMO、Gli1、Gli2的表达。结论:三氧化二砷联合伊曲康唑能抑制KG1a细胞增殖及诱导其凋亡,其机制可能与Hh信号通路受抑制,Caspase-3和BAX蛋白表达上调有关。对难治性AML进行三氧化二砷与伊曲康唑联合实验性治疗提供实验数据。
Objective: To explore the effect of arsenic trioxide combined with itraconazole on proliferation and apoptosis of KG1 a cells and its potential mechanism. Methods: The cell morphology was observed with Wrighe-Giemsa staining;cell survival rate was examined by CCK-8; and colony formation capacity was measured by methylcellulose colony formation test; the flow cytometry was used to analyse the cell apoptosis rate and cell cycle; the protein expressions of BCL-2, caspase-3,BAX,SMO,Gli1 and Gli2 were detected by Western-blot. Results: The arsenic trioxide and itraconazole alone both could inhibit the KG1a cell proliferation in dose-and time-dependent manner. In comparison between single and combined drug-treatment group,both the cell survival rate and the colony number of the single drug-treatment group were significantly lower( P〈0. 05),and the apoptosis rate was higher in the combined drug-treatment group. In the combined-treatment group,the protein expression of Caspase-3 and BAX was upregulated,while the protein expression of BCL-2,SMO,Gli1 and Gli2 was downregulated. Conclusion: Arsenic trioxide combined with itraconazole can inhibit the KG1a cell proliferation and induce apoptosis,which may be related with the inhibition of Hh signaling pathway and upregulation of both Caspase-3 and BAX protein expression,and provided experimental data of arsenic trioxide combined with itraconazole for the treatment of refractory AML.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2017年第4期1003-1010,共8页
Journal of Experimental Hematology
基金
2015年广东省江门市科技局科技计划立项项目(No.20150020)