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目的以小鼠白血病细胞系L1210为对象,探讨半胱天冬氨酸蛋白酶(Caspases)家族成员在二乙酰二脱水卫矛醇(diacetyldianhydrogalactitol,DADAG)诱导肿瘤细胞凋亡过程中的作用。方法MTT法测定DADAG对L1210细胞的杀伤作用;透射电镜和流式细胞术检测细胞凋亡;Western blot和Caspase-3试剂盒测定Caspase-3活性。结果与对照组相比,L1210细胞经不同浓度的DADAG(12.0、17.2、24.5、35.0或50.0 mg/L)处理72 h后,其存活率,分别降至72%、63%、46%、35%和20%。DADAG 50.0 mg/L处理组的凋亡率、Caspase-3活性和PARP裂解片段较对照组明显升高。Caspase-3抑制剂可部分阻断此过程,而Caspases抑制剂可完全阻断此过程。结论Caspases家族成员在DADAG诱导L1210细胞凋亡中发挥重要的作用。
Objective To investigate the role of caspases in the apoptosis of tumor cells induced by diacetyldianhydrogalactitol (DADAG) in mouse leukemia cell line L1210. Methods The killing effect of DADAG on L1210 cells was determined by MTT assay. Apoptosis was detected by transmission electron microscopy and flow cytometry. Caspase-3 activity was measured by Western blot and Caspase-3 kit. Results Compared with the control group, the survival rate of L1210 cells treated with different concentrations of DADAG (12.0,17.2,24.5,35.0 or 50.0 mg / L) for 72 h decreased to 72%, 63%, 46%, 35% % And 20%. The apoptosis rate, Caspase-3 activity and PARP cleavage fragment in DADAG 50.0 mg / L treatment group were significantly higher than those in control group. Caspase-3 inhibitors partially block this process, whereas Caspases inhibitors completely block this process. Conclusion Caspases family members play an important role in DADAG-induced L1210 cell apoptosis.