CAJ | 학술논문

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（ inhibi-tor），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。
목적：관찰미소RNA-16（microRNA-16，miR-16）대인백혈병세포K562향거핵세포계분화적영향，병초보탐색기중가능적궤제。방법：재K562세포중통과전염miR-16모의물（ mimics）혹miR-16억제물（ inhibi-tor），실시형광정량PCR검측miR-16적수평변화，통과류식세포술검측거핵세포계분화지표CD41、CD42b급CD61적표체。용Western blotting검측miR-16대하유백혈병암기인（ myeloblastosis oncogene，MYB）단백수평적영향，진이이용류식세포술검측miR-16시부통과영향MYB조공CD41、CD42b급CD61적표체。결과：전염miR-16-mimics가현저승고K562세포중적miR-16수평병촉진CD41、CD42b급CD61적표체（P＜0.05），이전염miR-16-inhibitor가명현억제K562세포중적miR-16수평동시강저CD41、CD42b급CD61적표체（ P＜0.05）；Western blotting증실miR-16가조공MYB단백수평，이침묵MYB가역전miR-16대CD41、CD42b급CD61표체적조공작용。결론：miR-16가통과조공MYB적표체，조절인백혈병세포K562향거핵세포계분화적능력。
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.