氟苯达唑对人急性髓系白血病HL-60细胞的增殖抑制作用研究

目的:研究氟苯达唑对人急性髓系白血病HL-60细胞增殖的抑制作用,明确氟苯达唑对HL-60细胞周期,凋亡发生的作用机制。方法:噻唑蓝法(MTT)检测氟苯达唑对人急性髓系白血病HL-60细胞的生长抑制作用,流式细胞术检测氟苯达唑对HL-60细胞周期,DNA片段化的影响,免疫印迹法检测Caspase, Raf, Bcl-2家族蛋白表达。结果:氟苯达唑抑制人急性髓系白血病HL-60细胞生长,HL-60细胞G2/M期增加,与阴性对照组相比,在一定的剂量和时间内,差别具有显著统计学意义;DNA片段化上升,0.25,0.5,1μM组与对照组相比差别具有显著统计学意义,促使Cleaved PARP,Cleaved-caspase 3,Cleaved-caspase 9蛋白表达量趋势增加;Bag-1和Bcl-2蛋白表达量降低;b-raf,c-raf磷酸化蛋白表达水平逐渐降低。结论:氟苯达唑通过诱导HL-60细胞阻滞于G2/M期,增加DNA片段化水平,激活Caspase, Raf, Bcl-2家族介导的凋亡相关通路抑制人急性髓系白血病HL-60细胞增殖,诱导人急性髓系白血病HL-60细胞发生凋亡而发挥抗肿瘤作用。

The Inhibitory Effect of Flubenazole on Proliferation of Human Acute Myeloid Leukemia HL-60 Cells

ABSTRACT Objective: To investigate the inhibitory effect of flubendazole on the proliferation of human acute myeloid leukemia HL-60 cells, and clarifying the mechanism of flubendazole on cell cycle and apoptosis of HL-60 cells. Methods: Methyl thiazolyl tetrazolium(MTT) assay was used to evaluatethe growth inhibition effects of flubendazole on human acute myeloid leukemia HL-60 cells. Flow cytometry was used to detect the effect of flubendazole on HL-60 cell cycle and DNA fragmentation. Western blotting was used to detect the expression of Caspase, Raf, and Bcl-2 family proteins. Results: Flubendazole inhibited the growth of human acute myeloid leukemia HL-60 cells. The G₂/M phase, the DNA fragmentation and the expression of Cleaved PARP, Cleaved-caspase 3, and Cleaved-caspase 9 of HL-60 cells increased. The expression of Bag-1 protein was decreased and the Bcl-2 was increased. The b-raf and the c-raf phosphorylation protein levels was inhibited by Flubendazole. Conclusion: The G₂/M phase of human acute myeloid leukemia HL-60 cells was arrested by flubendazole, and increasing DNA fragmentation level and activating Caspase, Raf, and Bcl-2 family apoptosis- related pathways which Inducing apoptosis in human acute myeloid leukemia HL-60 cells and taking anti-tumor effects.