摘要
目的 :研究国产氨肽酶 N抑制剂乌苯美司 ( U benim ex)对人白血病细胞的作用及其机理。方法 :应用MTT比色法观察 U benim ex对细胞的生长抑制作用 ,光学显微镜观察细胞形态结构的改变 ,DN A凝胶电泳、DNA片段原位末端标记及流式细胞仪 ( FCM)检测 ,分析细胞凋亡。结果 :1U benim ex明显抑制 H L6 0细胞的生长 ,半数抑制浓度 ( IC5 0 )为 13.0 3μg/m l;而 K 56 2细胞的敏感性较差。其抑制作用均呈剂量效应关系。 2典型的细胞形态改变 ,DN A片段化 ,DNA末端原位标记的检出及流式细胞仪结果 ,均证实 U benim ex能诱导白血病细胞的凋亡。 310μg/m l U benimex作用 12 h即可明显诱导 H L6 0细胞 DNA断裂 ,K56 2细胞的凋亡敏感性显著低于 H L6 0细胞 ;两者凋亡率均呈剂量和时间依赖性 ,经 U benim ex处理后 ,HL6 0细胞出现 G1期阻滞。结论 :U benim ex能抑制 K56 2 ,HL6 0细胞生长 ,诱导细胞凋亡是其机制之一。
Objective: To study the mechanism of inhibitory effect of Ubenimex on human leukemic cells. Me thods: K562 and HL60 cells were treated with Ubenimex at different concentrations, and the growth inhibition was analysed by MTT assay. Cell apoptosis was evaluated by light microscopy, agrose gel electrophoresis,TUNEL labeling method and flow cytometry(FCM) assay. Results: ①Treatment with Ubenimex remarkably inhibited the growth of HL60 cells,the IC 50 of Ubenimex for HL60 cells was 13.03 μg/ml. But K562 cells were less sensitive than HL60. Ubenimex inhibited the growth of HL60 and K562 cells in a dose dependent manner. ②Apoptpsis of leukemic cells was induced by Ubenimex, which was shown by the changes in morphology, DNA ladder on agrose gel, TUNEL labeling,typical peak before G1 phase of cell cycle and the positive of Annexin Ⅴ FITC on the cells membrane with FCM. ③Ubenimex induced apoptosis of K562 and HL60 cells in a dose and time dependent manner. ④The cell cycle analysis by FCM showed that the HL60 cells were blocked in G1 phase after treated by Ubenimex. Conclution: Ubenimex can efficiently induce apoptosis of HL60 and K562 cells, this may be one of the mechanisms for inhibiting effect of Ubenimex on leukemia.
出处
《浙江大学学报（医学版）》
CAS
CSCD
2002年第4期259-264,共6页
Journal of Zhejiang University(Medical Sciences)
