全反式维甲酸诱导HL-60细胞粒系分化中miRNA-26的表达变化

Alteration of miRNA-26 expression in the granulocytic differention of HL-60 cells induced by ATRA

目的探讨全反式维甲酸(ATRA)对HL-60细胞诱导分化的miRNA-26表达变化。方法采用全反式维甲酸(1μM)影响下不同时间的HL-60细胞为检测对象,显微镜下观察细胞形态学改变;流式细胞仪检测细胞表面分化标志CD15;Real-timePCR检测不同时段miRNA的表达变化。结果①随着药物诱导时间的延长,被诱导的HL-60细胞向粒系分化,并出现细胞核型的改变;②与对照组相比,ATRA作用3、4、5和6d后,流式细胞仪检测HL-60细胞的分化率分别为(52.39±3.56)%(,77.65±0.24)%,(83.90±1.30)%及(68.13±1.67)%,(P<0.001);③Real-timePCR检测miR-26A、miR-26B的表达水平显示,5d时miRNA表达水平显著上升,miR-26A、miR-26B的表达差异有统计学意义(P<0.05)。结论 miR-26A、miR-26B在ATRA诱导HL-60细胞粒系分化中表达显著升高,提示miR-26可能参与急性髓系白血病粒系分化调控的过程。

[Objective]To investigate the expression pattern of microRNA-26 （miR-26） in the differentiation of acute myeloblastic leukemia （AML） cell lines HL-60 induced by all-trans retinoic acid （ATRA）. [Methods]HL-60 cells stimulated by ATRA （1 μmol/L） capable of inducing differentiation for different time were used as subjects. The alterations in cell morphology were observed with Wright staining under microscope. The differentiation marker on the cell membrane surface was analyzed by using flow cytometry. The miRNAs expression was verified by using real-time fluorescence quantitative PCR. [Results]With drug-inducing time prolonged, the differentiation along the neutrophil lineage was proved and the nuclei morphology of the differentiation cells was changed. As compared with control group, the expression of differentiating rate which induced and differentiated by ATRA（1μmol/L） at 3 d, 4 d, 5 d and 6 d was （52.39 ± 3.56）%, （77.65 ± 0.24）%, （83.90 ± 1.30）%, （68.13 ± 1.67）%, （P 0.001）, respectively. The relative expressions of miRNA-26A and-26B were significantly up-regulated at 5 d, and showed remarkable difference （P 0.05）. [Conclusion]miR-26A and26B are highly expressed in the differentiation of HL-60 cells induced by ATRA, which indicated that miR-26 may be invovled in the regulation process of granulocytic differentiation, providing foundation for further study of the molecular mechanism.