Staurosporine诱导维A酸耐药早幼粒细胞白血病细胞株分化的研究

目的:研究蛋白激酶C抑制剂staurosporine对全反式维A酸(ATRA)耐药的急性早幼粒细胞白血病(APL)细胞株的诱导分化作用及其可能的机制。方法:以ATRA耐药的APL细胞株NB4-R1为模型,通过检测细胞表面分化抗原CD11b,并进行四唑氮蓝(NBT)还原实验和形态学观察,分析Staurosporine对NB4-R1细胞分化的影响;应用蛋白印迹法检测staurosporine对C/EBPβ、C/EBPε和PML-RARα蛋白表达水平的影响。结果:与对照组相比,staurosporine处理72 h后,细胞CD11b阳性率从(5.84±1.89)%上升到(65.90±2.00)%,差异有统计学意义(P

Study on staurosporine-induced differentiation in all-trans-retinoic acid-resistant APL cells

Objective： To study the differentiation inducing effect of PKC inhibitor staurosporine on all-trans-retinoic acid （ATRA） resistant acute promyeloeytic leukemia （APL） cells and its possible mechanisms. Methods： ATRA-resistant APL cell line NB4-R1 cells were used as the in-vitro model. The differentiation inducting effect of staurosporine on NB4- R1 cells was evaluated by detection of cell surface differentiation antigen CD1 lb, nitroblue-tetrazolium （NBT） reduction assay and cellular morphology observation. Meanwhile, the effect of staurosporine on protein expression levels of C/EBPβ, C/EBPeand PML-RARct was assayed by Western Blot. Results： Compared with control group, the percentage of CDllb positive cell in NB4-R1 cells treated with staurosporine for 72h increased significantly from （5.84±1.89） % to （65.90_＋2.00） % （P〈O.01）. Meanwhile, NBT reduction A~o value increased markedly from 0.033 0±0.000 5 to 0.103 0±0.014 9 （P〈O.O1）. Classical differentiated cells were observed. The protein expression levels of C/EBPI3 and C/EBPe were increased by staurosporine treatment. Conclusions： Staurosporine could induce the differentiation of ATRA-resistant APL cells.