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  

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

作　　者：蔡心颖 林东麒 陈清[2] 杨虹 郭江睿[4] 李暐[2] CAI Xin-ying;LIN Dong-qi;CHEN Qing;YANG Hong;GUO Jiang-rui;LI Wei(Clinical Medical Research Center,Shantou Central Hospital,Shantou,Guangdong,515000,China;School of Pharmacy,Fujian Medical University,Fuzhou,Fujian,350004,China;Department of Cardiothoracic Surgery,Second Affiliated Hospital of Shantou University Medical College,Shantou,Guangdong,515000,China;Fujian Institute of Hematology,Fujian Provincial Key Laboratory of Hematology,Union Hospital of Fujian Medical University,Fuzhou,Fujian,350004,China)

机构地区：[1]汕头市中心医院临床医学研究中心,广东汕头515000 [2]福建医科大学药学院,福建福州350004 [3]汕头大学医学院附属第二医院心胸外科,广东汕头515000 [4]福建省血液病研究所,福建省血液病学重点实验室,福建医科大学附属协和医院,福建福州350004

出　　处：《现代生物医学进展》2020年第4期648-653,686,共7页Progress in Modern Biomedicine

基　　金：福建省自然科学基金项目(2017J01531);福建省卫生健康中青年骨干人才培养项目(2018-ZQN-33);福建省青年科技人才创新项目(2017J05127)。

摘　　要：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。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 apoptosisrelated pathways which Inducing apoptosis in human acute myeloid leukemia HL-60 cells and taking anti-tumor effects.

关 键 词：氟苯达唑 急性髓系白血病 周期 凋亡 

分 类 号：R-33[医药卫生] R733.71