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目的:研究caspases家族成员在二乙酰二脱水卫矛醇(DADAG)诱导人白血病HL-60细胞凋亡中的作用.方法:MTT法观察DADAG的体外抗增殖作用;透射电镜、DNA梯形条带和流式细胞仪检测HL-60细胞凋亡;caspase-3检测试剂盒和Western blot法分析caspases家族成员.结果:DADAG明显抑制HL-60细胞增殖和诱导细胞凋亡.DADAG处理HL-60细胞24h后,caspase-3酶活性达峰值,同时聚腺苷二磷酸核糖聚合酶(PARP)、lamin B和DFF45蛋白开始出现断裂片段.Caspase-3抑制剂z-DEVD·fmk可部分逆转DADAG诱导的HL-60细胞凋亡,而caspases广谱抑制剂z-VAD·fmk可完全逆转此作用.结论:Caspases在DADAG诱导HL-60细胞凋亡中起重要作用,它们通过酶解底物PARP、DFF45和lamin B促进细胞凋亡.
AIM: To investigate the role of caspases in the apoptosis of human leukemia HL-60 cells induced by diacetyldianhydrogalactitol (DADAG) .Methods: MTT assay was used to observe the anti-proliferative effect of DADAG in vitro. Transmission electron microscopy, DNA ladder and The apoptosis of HL-60 cells was detected by flow cytometry, and caspases family members were analyzed by caspase-3 detection kit and Western blot.Results: DADAG significantly inhibited the proliferation and induced apoptosis of HL-60 cells.DADAG treatment of HL-60 cells 24h , The caspase-3 activity peaked and the cleaved fragments of poly (ADP-ribose polymerase) (PARP), lamin B and DFF45 proteins began to appear.Caspase-3 inhibitor z-DEVD · fmk partially reversed DADAG-induced HL -60, and caspases broad-spectrum inhibitor z-VAD · fmk can completely reverse this effect.Conclusion: Caspases play an important role in the apoptosis of HL-60 cells induced by DADAG.It can inhibit the apoptosis of HL-60 cells by digesting the substrates PARP, DFF45 and lamin B promotes apoptosis.