Embelin对HL-60细胞增殖、分化和凋亡的影响

Effect of embelin on proliferation, differentiation and apoptosis of HL-60 cells

目的 研究Embelin对HL-60细胞增殖、分化和凋亡的影响,并对其作用机制进行初步探讨.方法 用不同浓度的Embelin处理HL-60细胞,采用MTT法绘制生长曲线,通过Annexin V/PI 复染及JC-1染色,观察Embelin对HL-60细胞凋亡的影响 通过流式细胞术检测细胞表面分化抗原CD33、CD34、CD11b和CD14表达的变化.在对HL-60细胞研究的基础上,选择9例急性非淋巴细胞白血病（ANLL）患者的骨髓进行相应的研究.结果 Embelin对HL-60细胞具有增殖抑制作用,且作用呈时间和剂量依赖性.24 h的,IC50值为429.98 μmoL/L.33.97 μmol/L的Embelin作用HL-60细胞3 d,流式细胞术检测结果 显示CD11b、CD14阳性细胞率均升高（P值均〈0.01） 339.67 μmol/L Embelin作用于HL-60细胞后12、24及48 h的特异性凋亡率分别为（9.23 ±0.05）%、（25.86 ±0.30）%和（39.03±0.07）%,10.19、33.97、101.90、339.67及1019.02 μmol/L Embelin作用于HL-60细胞24 h后特异性凋亡率分别为（0.07±0.03）%、（7.43±0.30）%、（14.01±0.01）%、（25.52±0.03）%和（39.15±0.01）%,其诱导凋亡的作用呈时间及剂量依赖性.33.97 μmol/L的Embelin作用于ANLL 患者骨髓细胞3 d,3例M3患者骨髓细胞表面分化抗原出现显著性改变 339.67 μmol/L的Embelin作用患者骨髓细胞24 h,9例均发生凋亡.结论 亚细胞毒浓度的Embelin可诱导HL-60细胞向单核细胞分化 高浓度的Embelin对HL-60细胞具有增殖抑制及促进凋亡的作用.其促进凋亡的机制与线粒体凋亡途径有关.亚细胞毒浓度的Embelin对体外培养的M3患者骨髓细胞具有促进分化的作用 339.67 μmol/L的Embelin对ANLL骨髓细胞具有促进凋亡的作用.

Objective To study the effect of embelin on proliferation, differentiation and apoptosis of HL-60 cells and explore its possible mechanism. Methods Different concentration of embelin were used to treat HL-60 cells. Cell growth curve was analysed by MTT assay, cell apoptosis by Annexin V/PI double staining and JC-1 dye. The differentiation of HL-60 cells was evaluated by expression of CD33 , CD34, CD11 b and CD14. Bone marrow cells （BMC） from nine patients with acute nonlymphocytic leukemia（ ANML） were also studied. Results Embelin induced differentiation of HL-60 cells with significant increase of CD14 and CD1lb expression at 33. 97μ,mol/L for 3 days（ P 〈 0. 01 ）. Embelin induced apoptosis of HL-60 cells in a time- and dose-dependent manner, the apoptosis rates were （9. 23 0. 05 ） % , （ 25. 86 0. 30） % and （39.03 0. 07）% respectively at 339. 67 μmol/L of embelin for 12-, 24- and 48-hours treatment （P 〈 0.05） the apoptosis rates were （0. 07 0. 03）% , （7. 43 0. 30）% , （ 14. 01 0. 01 ）% , （25. 52 0.03）% and （39. 15 0.01）% respectively at 10. 19, 33.97, 101.90, 339.67 and 1019.02 μmol/L of embelin for 24-hours culture（P〈0.05）. Clusters of differentiation antigen on BMC from three acute promye-locytic leukemia patients showed significant changes at 33.97 μmol/L of embelin treatment for 3 days. Embelin induced apoptosis of BMCs from all the nine ANML patients at 33.97 μmol/L for 24 hour. Conclusion Embelin can inhibit proliferation and induce differentiation and apoptosis of HL-60 cells. The mechanism may be related to mitochondrial apoptosis pathway. Embelin at subtoxic concentration doesn＇t promote leukemia BMC differentiation, but at 339.67 μmol/L induces apoptosis of these cells.