土贝母苷甲对人HL-60髓性白血病细胞周期与凋亡的影响

Effects of TubeimosideI on Cell Cycle and Apoptosis of Human Myeloblastic Leukemia Cells(HL-60)

目的:研究土贝母苷甲(下称苷甲)对人髓性白血病细胞HL-60的细胞周期及凋亡的影响。方法:采用MTT(四甲基偶氮唑蓝)法检测苷甲对HL-60细胞生长的影响;形态学方法(荧光显微镜和透射电镜)、流式细胞仪和DNA琼脂糖凝胶电泳观察和分析在苷甲作用下HL-60细胞形态、DNA含量的变化和DNA断裂的情况;蛋白印迹免疫法检测细胞凋亡及周期相关基因bcl-2、cyclinB1表达的变化。结果:苷甲显著抑制HL-60细胞的生长,其抑制效果与浓度及时间呈依赖关系。15μmol·L-1苷甲作用24小时可将HL-60细胞阻滞于G2/M期,进而诱导其凋亡,凋亡细胞具有典型的凋亡形态特征,琼脂糖凝胶电泳可见明显的梯状区带。bcl-2表达无明显变化,cyclinB1表达降低。结论:苷甲对细胞周期起阻滞作用,并诱导其凋亡,其作用机制可能与cyclinB1表达降低有关。

Objective:To study the effects of tubeimoside I on cell cycle and apoptosis of human myeloblastic leukemia cells(HL-60).Methods:The growth inhibitory activity of tubeimo-side I was evaluated in HL-60cells with MTT assay.Cell cycle and the induction of apoptosis were evaluated by flow cytometry,fluorescent microscopy,electron microscopy and agarose gel electrophoresis.Western blot analysis was used to determine bcl-2and Cyclin B1expression.Re sults:Tubeimoside I displayed growth inhibitory activity against HL-60cells with IC 50 values ranging from13.0～30.7μmol·L -1 .HL-60cells underwent G 2 /M arrest within24hours of exposure to tubeimoside I,and had morphologic and biochemical evidences of apoptosis after72hours.Western blot analysis of tubeimoside I-treated cells revealed low CyclinB1expressing level con-sistent with G 2 /M arrest.Conclusion:Tubeimoside I has in vitro activity against HL-60cells,and this activity is associated with G 2 /M arrest,the induction of apoptosis,and low CyclinB1ex-pressing level.These findings suggest that further evaluation of the activity of tubeimoside I in HL-60cells is reasonable and that the pharmacologic modulation of cell cycle arrest and apopto-sis pathways deserves further study as a potential anticancer strategy.