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目的:研究MEK-ERK信号通路在全反式维A酸(all-trans retinoic acid,ATRA)诱导急性早幼粒细胞白血病(acute promyelocytic leukemia,APL)细胞分化中的作用及其可能的机制。方法:采用APL细胞株NB4作为体外模型,以细胞表面分化抗原CD11b、四唑氮蓝(NBT)还原实验和形态学观察评估细胞分化;应用蛋白印迹法研究细胞MEK和ERK的活化状态及PU.1、C/EBPβ、C/EBPε和PML-RARα的蛋白含量。结果 :MEK-ERK信号通路在ATRA处理早期即活化,并维持48 h。抑制MEK活性,可使ATRA诱导的CD11b阳性率从(87.50±4.16)%下降到(38.01±2.79)%,NBT A540值从0.507±0.009下降到0.250±0.010,差异均有统计学意义(P<0.01和P<0.000 1);且大部分细胞形态回复到原始细胞;同时,ATRA诱导的PU.1、C/EBPβ和C/EBPε蛋白表达上调也受阻。结论:ATRA可通过MEKERK信号通路调控PU.1、C/EBPβ和C/EBPε蛋白表达,诱导APL细胞分化。
AIM: To investigate the role of MEK-ERK signaling pathway in the differentiation of acute promyelocytic leukemia (APL) cells induced by all-trans retinoic acid (ATRA) and its possible mechanism. Methods: APL cell line NB4 was used as an in vitro model to evaluate cell differentiation by cell surface differentiation antigen CD11b, NBT reduction assay and morphological observation. The activation status and PU of MEK and ERK were studied by Western blotting. 1, C / EBPβ, C / EBPε and PML-RARα. Results: The MEK-ERK signaling pathway activated early in ATRA treatment and maintained for 48 h. The inhibition of MEK activity decreased the percentage of CD11b-positive cells from (87.50 ± 4.16)% to (38.01 ± 2.79)%, and NBT A540 decreased from 0.507 ± 0.009 to 0.250 ± 0.010 (P < 0.01 and P <0.0001). Most of the cell morphology returned to the original cells. At the same time, the upregulation of ATRA-induced PU.1, C / EBPβ and C / EBPε proteins was blocked. CONCLUSION: ATRA can regulate the expression of PU.1, C / EBPβ and C / EBPε through MEKERK signaling pathway and induce APL cell differentiation.