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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 effect and mechanism of domestic aminopeptidase N inhibitor U benim ex on human leukemia cells. Methods: The growth inhibition of U benim ex was observed by MTT colorimetric assay. The morphological changes of cells were observed by light microscopy. The DNA was detected by gel electrophoresis (DN A), DNA end labeling in situ and flow cytometry (FCM) Apoptosis. Results: 1U benim ex significantly inhibited the growth of HL-60 cells, and the IC50 was 13.0 3μg / ml; however, the sensitivity of K 562 cells was poor. The inhibitory effects were dose-response relationship. 2 Typical morphological changes of cells, DN A fragmentation, detection of DNA end-in-situ markers and flow cytometry results confirm that U benim ex induces apoptosis in leukemic cells. The DNA fragmentation of HL-60 cells was induced by 310μg / ml U benimex for 12 h, and the apoptosis sensitivity of HL-60 cells was significantly lower than that of HL-60 cells. The apoptosis rates of both were dose-and time-dependent, After treated with benim ex, HL60 cells showed G1 arrest. Conclusion: U benim ex can inhibit the growth of K56 2 and HL60 cells and induce apoptosis.