微小RNA-16对K562细胞向巨核细胞系分化的影响

Effect of miR-16 on megakaryocytic differentiation of K562 cells

目的:观察微小RNA-16(microRNA-16,miR-16)对人白血病细胞K562向巨核细胞系分化的影响,并初步探索其中可能的机制。方法:在K562细胞中通过转染miR-16模拟物(mimics)或miR-16抑制物(inhibitor),实时荧光定量PCR检测miR-16的水平变化,通过流式细胞术检测巨核细胞系分化指标CD41、CD42b及CD61的表达。用Western blotting检测miR-16对下游白血病癌基因(myeloblastosis oncogene,MYB)蛋白水平的影响,进而利用流式细胞术检测miR-16是否通过影响MYB调控CD41、CD42b及CD61的表达。结果:转染miR-16-mimics可显著升高K562细胞中的miR-16水平并促进CD41、CD42b及CD61的表达(P<0.05),而转染miR-16-inhibitor可明显抑制K562细胞中的miR-16水平同时降低CD41、CD42b及CD61的表达(P<0.05);Western blotting证实miR-16可调控MYB蛋白水平,而沉默MYB可逆转miR-16对CD41、CD42b及CD61表达的调控作用。结论:miR-16可通过调控MYB的表达,调节人白血病细胞K562向巨核细胞系分化的能力。

AIM： To observe the effect of microRNA-16 （miR-16） on the megakaryocytic differentiation of K562 cells, and to explore the potential mechanism.METHODS：miR-16 was over-expressed or silenced by transfection with miR-16 mimics or inhibitor in K562 cells.The level of miR-16 was detected by real-time PCR.The expression of CD41, CD42b and CD61, as megakaryocytic differentiation markers, was detected by flow cytometry.The effect of miR-16 on the expression of myeloblastosis oncogene （ MYB） was measured by Western blotting, and flow cytometry was performed to confirm whether the effect of miR-16 on expression of CD41, CD42b and CD61 was mediated by MYB.RESULTS：Transfection with miR-16 mimics dramatically elevated the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells.Transfection with miR-16 inhibitor decreased the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells （P〈0.05）.The expression of MYB was regulated by miR-16, and MYB silencing reversed the regulation of CD41, CD42b and CD61 induced by miR-16.CONCLUSION：miR-16 regulates the megakaryocytic dif-ferentiation of K562 cells by targeting MYB.