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EVI1 Is Expressed in Megakaryocyte Cell Lineage and Enforced Expression of EVI1 in UT-7/GM Cells Induces Megakaryocyte Differentiation

https://doi.org/10.1006/bbrc.2002.6693Get rights and content

Abstract

3q21q26 syndrome, an acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) with chromosomal translocations or inversions between the bands 3q21 and 3q26, is frequently associated with dysmegakaryocytopoiesis and increased platelet counts at the initial diagnosis. Since the EVI1 gene at 3q26 is transcriptionally activated in 3q21q26 syndrome, we assessed the role of EVI1 gene expression in the abnormal megakaryocytic differentiation in 3q21q26 syndrome. RT-PCR analysis of various types of hematopoietic cells revealed that the EVI1 gene is expressed specifically in CD34+ cells, megakaryocytes, and platelets. UT-7 is a human immature megakaryoblastic leukemia cell line with dependence for the growth on granulocyte–macrophage colony-stimulating factor (GM-CSF) (designated at UT-7/GM) and with a differentiation capacity to erythroid (UT-7/EPO) and megakaryocytic lineages (UT-7/TPO) by erythropoietin (EPO) and thrombopoietin (TPO), respectively. Among three UT-7 sublines, UT-7/GM, UT-7/EPO, and UT-7/TPO, expression of the EVI1 gene was detected at low levels in UT-7/GM and UT-7/EPO cells, but was detected at a higher level in UT-7/TPO cells. When UT-7/GM cells were cultured with TPO, the level of EVI1 expression was increased, along with increased numbers of polynuclear megakaryocytes and expression of the platelet factor 4 (PF-4) gene. Furthermore, forced expression of the EVI1 gene in UT-7/GM cells changed their morphology to polynuclear megakaryocytes, stopped their growth, and induced cell death within a month. These data indicate that expression of the EVI1 gene is involved in progression of megakaryocytic differentiation and, thus, the dysmegakaryocytopoiesis in 3q21q26 syndrome could be partly due to an enhanced differentiation capacity of leukemia cells and/or megakaryocytes by constitutive expression of the EVI1 gene.

References (41)

  • M.H. Prandini et al.

    The tissue-specific transcriptional regulation of the megakaryocytic glycoprotein IIb gene is controlled by interactions between a repressor and positive cis-acting elements

    Blood

    (1996)
  • S. Deveaux et al.

    Analysis of the thrombopoietin receptor (MPL) promoter implicates GATA and Ets proteins in the coregulation of megakaryocyte-specific genes

    Blood

    (1996)
  • T. Tanaka et al.

    EVI1 raises AP-1 activity and stimulates c-fos promoter transactivation with dependence on the second zinc finger domain

    J. Biol. Chem.

    (1994)
  • K. Saitoh et al.

    Fluorescence in situ hybridization of progenitor cells obtained by fluorescence-activated cell sorting for the detection of cells affected by chromosome abnormality trisomy 8 in patients with myelodysplastic syndromes

    Blood

    (1998)
  • N. Takahashi et al.

    Lineage involvement of stem cells bearing the Philadelphia chromosome in chronic myeloid leukemia in the chronic phase as shown by a combination of fluorescence-activated cell sorting and fluorescence in situ hybridization

    Blood

    (1998)
  • M.L. Mucenski et al.

    Identification of a common ecotropic viral integration site, EVI1, in the DNA of AKXD murine myeloid tumors

    Mol. Cell. Biol.

    (1988)
  • K. Morishita et al.

    Activation of EVI1 gene expression in human acute myelogenous leukemias by translocations spanning 300–400 kilobases on chromosome band 3q26

    Proc. Natl. Acad. Sci. USA

    (1992)
  • K. Ohyashiki et al.

    EVI1 expression associated with a 3q26 anomaly in a leukemia cell line derived from the blast crisis of chronic myeloid leukemia

    Leukemia

    (1994)
  • K. Mitani et al.

    Generation of the AML1-EVI1 fusion gene in the t(3;21)(q26;q22) causes blastic crisis in chronic myelocytic leukemia

    EMBO J.

    (1994)
  • G. Nucifora et al.

    Consistent intergenic splicing and production of multiple transcripts between AML1 at 21q22 and unrelated genes at 3q26 in (3;21)(q26;q22) translocations

    Proc. Natl. Acad. Sci. USA

    (1994)
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    Work was supported in part by Grants-in-Aid from the Ministry of Health and Welfare.

    1

    To whom correspondence and reprint requests should be addressed at Department of Biochemistry, Miyazaki Medical College, 5200 Kihara, Miyazaki-Gun, Miyazaki-Ken 889-1692, Japan. Fax: 81-985-85-2401. E-mail: [email protected].

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