尿多酸肽诱导急性早幼粒细胞白血病细胞株凋亡的实验研究

摘要

目的 探讨尿多酸肽(CDA-Ⅱ)对急性早幼粒细胞白血病(acute promyelocytic leukemia,APL)细胞NB4及耐维甲酸的NB4-R1细胞的凋亡作用.方法 以NB4和NB4-R1细胞作为体外模型,观察CDA-Ⅱ作用前后细胞的形态和生长增殖情况.流式细胞术检测细胞凋亡特征.Western印迹法检测凋亡调控蛋白蛋白激酶C(PKCδ)及Caspase-3的水解活化.结果 CDA-Ⅱ作用于NB4和NB4-R1细胞后,细胞生长受到抑制；形态上观察到分化和凋亡现象；Annexin V-PI法发现,用药24 h后NB4和NB4-R1细胞凋亡比例分别上升至(58.7±3.1)％和(87.8±0.5)％；PKCδ和Caspase-3水解活化.结论 CDA-Ⅱ作用于细胞后,水解活化Caspase-3,裂解PKCδ生成活性片段,诱导细胞凋亡.%Objective To investigate the apoptosis-inducing effect of Uroacitides(CDA- II) on acute promyelocytic leukemia (APL) cell line NB4 and NB4-R1 (retinoic acid resistant). Methods The cultured NB4 and NB4-R1 cells were treated with different concentrations of CDA- II. The changes of cell morphology were observed; cell proliferation and growth inhibition was measured by cell counting; apoptosis was examined by flow cytometry; the activation of protein kinase C (PKC8) and Caspase-3 was detected by Western blotting. Results After treated by CDA- II the growth of NB4 and NB4-R1 cells was inhibited; morphological differentiation and apoptosis were observed. Annexin V-PI method showed that cell apoptosis rates were increased to (58.7 ± 3.1)% and (87.8 + 0.5)% for NB4 and NB4-R1 cells respectively after 24 h of treatment, and PKC8 and Caspase-3 were activated. Conclusion CDA- E can induce apoptosis of NB4 and NB4-R1 cells by activating the protein Caspase-3 and PKC5.