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目的 研究二乙酰二脱水卫矛醇 (DADAG)诱导人白血病HL 60细胞凋亡及其机理。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 ;Westernblotting法和caspase 3检测试剂盒分析DADAG诱导HL 60细胞凋亡与Bcl 2家族成员和caspase 3的关系。结果 DADAG明显抑制HL 60细胞增殖和诱导细胞发生凋亡。 8μg·mL- 1 DADAG处理HL 60细胞不同时间后 ,Bcl XL 蛋白水平呈时间依赖性地下降 ,而Bad蛋白水平上调。DADAG处理HL 60细胞 2 4h后 ,caspase 3酶活性达峰值。Caspase 3抑制剂z DEVD .fmk可部分逆转DADAG诱导HL 60细胞凋亡的作用 ,而caspases广谱抑制剂z VAD .fmk可完全逆转此作用。结论 DADAG诱导HL 60细胞凋亡依赖caspase 3途径的激活 ,而caspase 3的激活可能与Bcl 2家族成员密切相关
Objective To study the apoptosis induced by diacetyldianhydrogalactitol (DADAG) and its mechanism in human leukemia HL 60 cells. Methods MTT assay was used to observe the anti-proliferative effect of DADAG in vitro. The apoptosis of HL-60 cells was detected by transmission electron microscopy, DNA ladder and flow cytometry. The apoptosis of HL 60 cells induced by DADAG was analyzed by Western blotting and caspase 3 assay. Relationship between members and caspase 3. Results DADAG significantly inhibited HL 60 cell proliferation and induced cell apoptosis. After treated with 8μg · mL-1 DADAG for different time, the level of Bcl-XL protein decreased in a time-dependent manner, while the level of Bad protein was up-regulated. After treated with DADAG for 24 h, the caspase 3 activity peaked. Caspase 3 inhibitor z DEVD .fmk partially reversed the effect of DADAG-induced HL-60 cell apoptosis, whereas caspases-based broad-spectrum inhibitor z VAD .fmk completely reversed this effect. Conclusion DADAG induces the apoptosis of HL-60 cells dependent on the activation of caspase 3 pathway, while the activation of caspase 3 may be closely related to the Bcl-2 family members