异十三基二乙胺诱导HL-60细胞凋亡及其作用机制

目的探讨异十三基二乙胺(D-108)体外诱导人早幼粒白血病HL-60细胞凋亡及其作用机制。方法应用生长曲线法检测D-108对HL-60细胞生长抑制作用;相差显微镜观察药物对HL-60细胞促凋亡形态学变化;流式细胞仪分析细胞周期改变;DNA琼脂糖凝胶电泳法检测凋亡"梯状"DNA;比色法测定caspase-3,8,9酶活性:结果D-108显著抑制HL-60细胞生长,作用呈浓度及时间依赖性。HL-60细胞经D-108处理后,呈典型凋亡形态学改变,出现亚二倍体细胞峰及DNA梯状条带。caspase-3,9酶活性显著升高(P<0.01),而对caspase-8酶活性无明显影响。结论D-108在体外能有效诱导HL-60细胞凋亡,其作用机制可能通过线粒体凋亡途径。

Inducement and mechanism of allotri-tridecyl diethylamine on apoptosis of HL-60 cells

AIM To investigate the inducement and mechanism of allotri-tridecyl diethylamine(D-108) on the apoptosis of HL-60 cells in vitro.METHODS The inhibitory effect of D-108 on HL-60 cells was determined with growth curve method.Apoptosis morphological transformation of HL-60 cells induced by D-108 was detected through the phase contrast microscopy,alteration of cell cycle with flow cytometer and DNA ladder by agarose gel electrophoresis.The enzymatic activity of caspase-3,caspase-8 and caspase-9 were assayed by colorimetry.RESULTS The growth of HL-60 cells was significantly inhibited by D-108 with time-and dose- dependent,together with demonstrating typical apoptosis morphological transformation,sub-G_1 apoptotic peaks, and zones of DNA ladder.At the same time,the enzymatic activities of caspase-3 and caspase-9 were significantly higher,but no obvious change in caspase-8.CONCLUSION D-108 can induce the apoptosis of HL-60 cells effectively in vitro with the mechanism probably through mitochondria apoptosis pathway.