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目的 探讨一种新型的DNA拓扑酶 Ⅱ抑制剂磷烯菌素 (FST)对人急性粒细胞白血病细胞株HL 60的杀伤机理。方法 采用流式细胞术 (FCM )、TdT联合FCM及LadderDNA三种方法研究FST对HL 60细胞凋亡的诱导作用及其与增殖周期的关系。结果 :FCM (乙醇抽提法 )显示 :1 0 μmolFST作用 6h未引起HL 60凋亡 ,1 0 0 μmol/L时出现明显“亚G1 峰” ,但1 0 0 0 μmol/L诱导时 ,却不出现凋亡而呈“冰冻”现象。TdT联合FCM表明 :1 0 μmol/LFST作用 6h ,即有明显细胞凋亡 ,且凋亡发生在整个细胞周期。LadderDNA测定提示 :1 0 0 μmol/LFST作用HL 60细胞 6h ,其DNA凝胶电泳出现典型梯状图谱。结论 1 FST诱导HL 60细胞死亡具有异质性 :低剂量触发凋亡 ,高剂量诱导坏死。 2 TdT联合FCM是一种灵敏的凋亡检测手段 ,它能同时测定细胞DNA含量及断裂 ,故能用以研究细胞凋亡及其与增殖周期的关系 ;3 FST诱导的凋亡似乎没有细胞周期特异性
Objective To investigate the mechanism of killing of human acute myeloid leukemia cell line HL 60 by a new type of DNA topoisomerase inhibitor (FST). Methods Flow cytometry (FCM), TdT combined with FCM and Ladder DNA were used to study the effect of FST on HL-60 cell apoptosis and its relationship with the proliferation cycle. Results: The results of FCM showed that HL 60 apoptosis was not induced by 10 μmol FST for 6 h and “sub-G1 peak” was observed at 100 μmol / L, but when induced by 100 μmol / L, Apoptosis did not appear in the “frozen” phenomenon. TdT combined with FCM showed that: 1 0 μmol / LFST role 6h, that is, significant apoptosis, and apoptosis occurred in the entire cell cycle. The results of LadderDNA showed that HL 60 cells were treated with 100 μmol / L LST for 6 h, and typical DNA ladder gel electrophoresis showed a typical ladder pattern. Conclusions 1 FST induces heterogeneity of HL 60 cell death: low dose triggers apoptosis and high dose induces necrosis. 2 TdT combined with FCM is a sensitive apoptosis detection means that can simultaneously determine the cell DNA content and fragmentation, it can be used to study the relationship between apoptosis and proliferation cycle; 3 FST-induced apoptosis seems to be no cell cycle Specificity