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Transcription factor NF-E2 is essential for the polyploidization of a human megakaryoblastic cell line, Meg-J
S Kobayashi1, M Teramura, K Ito
1Department of Hematology, Tokyo Women's Medical College, Japan.
Abstract:
Transcription factors regulating the process of megakaryocyte development remain largely unclarified. To clarify them further, we used a human megakaryoblastic cell line, Meg-J, which showed prominant polyploidization and augmented platelet glycoprotein (GP) Ib expression after incubation with thrombopoietin (TPO, c-mpl ligand) and K252a (an indolocarbasole derivative). Under these conditions, we analyzed the expression of the transcription factors and observed that the expression of NF-E2 p45, but not those of GATA-1, GATA-2, Tal-1/SCL, Evi-1, and MafK, was increased after TPO and K252a stimulation. Gel-shift assay confirmed the enhanced binding activity to the NF-E2 site. The abolishment of NF-E2 p45 with NF-E2 antisense oligomers inhibited TPO plus K252a-induced polyploidization. These findings suggest that NF-E2 p45 is essential for the polyploidization of megakaryocytic cells.
Insights
Nuclear factor-erythroid 2 p45 (NF-E2 p45) is crucial for megakaryocyte polyploidization. This study demonstrates NF-E2 p45
Area of Science:
- Hematology
- Molecular Biology
- Cell Biology
Background:
- Megakaryocyte development and polyploidization are critical for platelet production.
- The transcription factors governing megakaryocyte differentiation are not fully understood.
Purpose of the Study:
- To investigate the role of transcription factors in megakaryocyte polyploidization.
- To identify key regulators of megakaryocyte development.
Main Methods:
- Utilized the human megakaryoblastic cell line Meg-J.
- Stimulated cells with thrombopoietin (TPO) and K252a.
- Analyzed transcription factor expression using gel-shift assays and antisense oligomers.
Main Results:
- Thrombopoietin (TPO) and K252a stimulation increased Nuclear factor-erythroid 2 p45 (NF-E2 p45) expression and binding activity.
- Expression of GATA-1, GATA-2, Tal-1/SCL, Evi-1, and MafK remained unchanged.
- Inhibition of NF-E2 p45 using antisense oligomers blocked TPO and K252a-induced polyploidization.
Conclusions:
- Nuclear factor-erythroid 2 p45 (NF-E2 p45) plays an essential role in megakaryocytic cell polyploidization.
- NF-E2 p45 is a key regulator in thrombopoietin-induced megakaryocyte differentiation.