红细胞膜蛋白是成熟红细胞行使功能的重要元件，但膜蛋白生成调控机制尚不明确。申请人首次发现红细胞重要结构和功能蛋白α-spectrin、Band 3在红系终末分化阶段显著增加；Deep sequence结果显示两个膜蛋白分子mRNA相对恒定；miRNAs array 结果显示miR-150呈阶段性下降20多倍；生物信息学表明miR-150靶向α-spectrin和Band 3 mRNA的3′UTR，在Hela细胞中荧光素酶实验发现miR-150与这两个分子mRNA 的3′UTR有结合。申请人据此提出在造血系统分化中发挥关键节点作用的miR-150在翻译后水平调控α-spectrin和Band3表达并影响红系终末分化；进而探讨anti-miR-150协同红系诱导分化剂诱导红白血病K562细胞向红系分化的作用和机制。本研究将揭示miR-150在红系发育中的功能，为白血病治疗提供新策略。

Formation of erythrocyte membrane skeleton is a critical part of erythropoiesis because this enables erythrocyte to circulate in capillary to fulfill its oxygen delivery function. However, the regulatory mechanism for the formation of this unique membrane skeleton is not well studied. We have recently discovered that the formation of erythrocyte membrane is characterized by the increased expression of major red cell membrane proteins such as α-spectrin and Band 3 during terminal erythrocyte differentiation. However, deep sequencing analysis revealed that the transcripts for these two proteins are stabilization. Furthermore, miRNA array analysis showed that miR-150 decreased more than 20 folders during terminal erythrocyte differentiation and bioinformatics software found miR-150 targets the 3′UTR of α-spectrin and Band 3 mRNA. Luciferase-based reporter assay documented the direct binding of miR-150 to the 3′UTR of these two gene mRNA in Hela cells. Based on these findings, we hypothesize that miR-150 (which plays an important role in lineage commitment during hematopoiesis) may modulate terminal erythroid differentiation by directly targeting α-spectrin and Band 3 expression. In the application, we will explore the effect of anti-miRNA-150 combine with erythropoietin on K562 cells differentiation and its underlying mechanism. The successful accomplishment of our proposed studies will reveal the role of miR-150 in erythropoiesis and may provide insights into the diagnosing and treating of various hemolytic anemias.