Thrombopoietic effects of CCAAT/enhancer-binding protein β on the early-stage differentiation of megakaryocytes

Abstract

CCAAT/enhancer-binding protein β (C/EBPβ) is a transcription factor that is involved in adipocytic and monocytic differentiation. However, the physiological role of C/EBPβ in megakaryocytes (MKs) is not clear. In this study, we investigated the effects of C/EBPβ on the early-stage differentiation of MKs, and explored the potential mechanisms of action. We established a cytosine arabinoside-induced thrombocytopenia mouse model using C57BL/6 mice. In the thrombocytopenia mice, the platelet count was found to be decreased, and the mRNA and protein expression levels of C/EBPβ in MKs were also reduced. Furthermore, the maturation of Dami (MKs cell line) cells was induced by phorbol 12-myristate 13-acetate. When C/EBPβ was silenced in Dami cells by transfection using C/EBPβ-small interfering RNA, the expression of MKs-specific markers CD41 and CD62P, was dramatically decreased, resulting in morphological changes and differentiation retardation in low ploidy, which were evaluated using flow cytometry, real-time polymerase chain reaction, western blot, and confocal microscopy. The mitogen activated protein kinase-extracellular signal-regulated kinase signaling pathway was found to be required for the differentiation of MKs; knockdown of C/EBPβ in MEK/ERK1/2 pathway attenuated MKs differentiation. Overexpression of C/EBPβ in MEK/ERK1/2 pathway inhibited by U0126 did not promote MKs differentiation. To the best of our knowledge, C/EBPβ plays an important role in MKs differentiation and polyploidy cell cycle control. Taken together, C/EBPβ may have thrombopoietic effects in the differentiation of MKs, and may assist in the development of treatments for various disorders.

Keywords: CCAAT/enhancer-binding protein β; CD41; CD62P; DNA ploidy; Megakaryocyte differentiation; Thrombopoiesis.