miR-125b modulates megakaryocyte maturation by targeting the cell-cycle inhibitor p19(INK4D).
miR-125b modulates megakaryocyte maturation by targeting the cell-cycle inhibitor p19(INK4D).
复制标题
miR-125b 通过靶向细胞周期抑制剂 p19(INK4D) 来调节巨核细胞成熟。
DOI：
10.1038/cddis.2016.288
复制
发表时间：
2016-10-20
影响因子：
9
通讯作者：
Pei X
中科院分区：
文献类型：
--
作者：
Qu M;Fang F;Zou X;Zeng Q;Fan Z;Chen L;Yue W;Xie X;Pei X
对巨核细胞成熟机制的深入了解将有助于血小板的体外培养及其临床应用。一种微小RNA，miR-125 b，已被认为在巨核细胞-红系祖细胞的自我更新和血小板生成中具有重要作用。然而，miR-125 b对造血干细胞自我更新也至关重要。因此，miR-125 b的功能和调节巨核细胞生成的复杂信号通路仍有待阐明。本研究对巨核细胞分化过程中miR-125 b的内源性表达进行了细致的检测。因此，造血干细胞的分化需要miR-125 b的下调，而巨核细胞的确定和成熟与miR-125 b的积累同步。miR-125 b的过表达促进K562和UT-7细胞的巨核细胞分化。此外，原代细胞中miR-125 b的阶段特异性过表达表明，miR-125 b介导了巨核细胞确定后巨核细胞分化的增强，在该阶段，巨核细胞对于造血祖细胞标志物CD 34的表达是阴性的。在巨核细胞生成过程中，miR-125 b靶点的鉴定主要集中在细胞周期的负调控因子上，因为G1/S期的转变与巨核细胞多倍化有关。实时荧光定量PCR、western blot和荧光素酶报告基因检测结果表明p19 INK 4D是miR-125 b的直接靶点。在巨核细胞诱导的K562细胞、UT-7细胞和CD 61+前巨核细胞中使用小干扰RNA（siRNA）敲低P19 INK 4D导致S期进展和增加的多倍性，以及改善的巨核细胞分化，类似于miR-125 b过表达的作用。P19 INK 4D过表达逆转了这些效应，如巨核细胞标志物表达减少、G1期阻滞和多倍性降低所示。miR-125 b下调细胞中的P19 INK 4D敲低或miR-125 b上调细胞中的p19 INK 4D过表达挽救了miR-125 b的作用。综上所述，这些发现表明，miR-125 b的表达正调控巨核细胞的发展，因为巨核细胞的决定的初始阶段，和p19 INK 4D是在巨核细胞多倍化的发病过程中的miR-125 b活性的关键介质之一。
A better understanding of the mechanisms involved in megakaryocyte maturation will facilitate the generation of platelets in vitro and their clinical applications. A microRNA, miR-125b, has been suggested to have important roles in the self-renewal of megakaryocyte-erythroid progenitors and in platelet generation. However, miR-125b is also critical for hematopoietic stem cell self-renewal. Thus, the function of miR-125b and the complex signaling pathways regulating megakaryopoiesis remain to be elucidated. In this study, an attentive examination of the endogenous expression of miR-125b during megakaryocyte differentiation was performed. Accordingly, the differentiation of hematopoietic stem cells requires the downregulation of miR-125b, whereas megakaryocyte determination and maturation synchronize with miR-125b accumulation. The overexpression of miR-125b improves megakaryocytic differentiation of K562 and UT-7 cells. Furthermore, stage-specific overexpression of miR-125b in primary cells demonstrates that miR-125b mediates an enhancement of megakaryocytic differentiation after megakaryocyte determination, the stage at which megakaryocytes are negative for the expression of the hematopoietic progenitor marker CD34. The identification of miR-125b targets during megakaryopoiesis was focused on negative regulators of cell cycle because the transition of the G1/S phase has been associated with megakaryocyte polyploidization. Real-time PCR, western blot and luciferase reporter assay reveal that p19INK4D is a direct target of miR-125b. P19INK4D knockdown using small interfering RNA (siRNA) in megakaryocyte-induced K562 cells, UT-7 cells and CD61+ promegakaryocytes results in S-phase progression and increased polyploidy, as well as improved megakaryocyte differentiation, similarly to the effects of miR-125b overexpression. P19INK4D overexpression reverses these effects, as indicated by reduced expression of megakaryocyte markers, G1-phase arrest and polyploidy decrease. P19INK4D knockdown in miR-125b downregulated cells or p19INK4D overexpression in miR-125b upregulated cells rescued the effect of miR-125b. Taken together, these findings suggest that miR-125b expression positively regulates megakaryocyte development since the initial phases of megakaryocyte determination, and p19INK4D is one of the key mediators of miR-125b activity during the onset of megakaryocyte polyploidization.
登录
查看更多内容
DOI：
10.1073/pnas.1016218107
发表时间：
2010-12-14
影响因子：
11.1
作者：
Ooi, A. G. Lisa;Sahoo, Debashis;Park, Christopher Y.
通讯作者：
Park, Christopher Y.
影响因子：
10.4
作者：
Chang, Y.;Bluteau, D.;Vainchenker, W.
通讯作者：
Vainchenker, W.
影响因子：
16
作者：
Lal A;Navarro F;Maher CA;Maliszewski LE;Yan N;O'Day E;Chowdhury D;Dykxhoorn DM;Tsai P;Hofmann O;Becker KG;Gorospe M;Hide W;Lieberman J
通讯作者：
Lieberman J
影响因子：
10.5
作者：
Klusmann, Jan-Henning;Li, Zhe;Reinhardt, Dirk
通讯作者：
Reinhardt, Dirk
影响因子：
20.3
作者：
Teofili, L;Martini, M;Larocca, LM
通讯作者：
Larocca, LM