AP-1/c-jun and c-myc regulation during megakaryocytic differentiation of a human bi-potential growth-factor-dependent cell line

H. G. Derigs, D. A. Morgan, R. Hoffman, S. L. Litz, Edward Srour, J. E. Brandt, H. Boswell

Research output: Contribution to journalArticle

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Abstract

Terminal megakaryocytic development is characterized by nuclear poly ploidization, appearance of specific granules, and enhanced expression of membrane platelet glycoproteins. We utilized a human GM-CSF-dependent cell line, MB-02, which is capable of under going megakaryocytic differentiation, to examine the molecular events underlying this process. The responses of MB-02 to the protein kinase C (PKC) agonist, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) were examined. GM-CSF dependent proliferation of MB-02, as measured by 3H-thymidine uptake, was greater than 95% inhibited by TPA (16 nM), but was not affected by the inactive stereoisomer, 4-αphorbol-12,13-didecanoate (4-αPDD). Transient exposure of cells to GM-CSF after growth factor deprivation led to rapid, high-level expression of normal-sized c-myc mRN A transcripts above baseline. C-myc expression was turned off by TPA (16 nM) stimulation of cells within 2-4 h. This TPA-mediated effect likely occurred at the transcriptional level since the half life of c-myc mRN A induced by GM-CSF was less than 30 min. Treatment of cells with TPA was associated with induction of c-jun and junB mRN A within 1-4 h. The protein products of these transcription factors are known to be part of the transcription factor complex Activator protein 1 (AP-1). Indeed, our data prove a rapid induction of AP-1 protein after TPA stimulation, as shown by mobility shift assays. In addition, TPA treatment resulted in expression of platelet surface glycoprotein IIb/IIIa complex (gpIIb/IIIa). These studies suggest a link between PKC stimulation by TPA and AP-1 activation with downregulation of c-myc transcription on a molecular level. At the cellular level, PKC activation was related to the acquisition of several features of the megakaryocyte development programme associated with the switch from cell proliferation to maturation.

Original languageEnglish
Pages (from-to)24-30
Number of pages7
JournalPlatelets
Volume6
Issue number1
DOIs
StatePublished - 1995

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Transcription Factor AP-1
Intercellular Signaling Peptides and Proteins
Acetates
Cell Line
Granulocyte-Macrophage Colony-Stimulating Factor
Protein Kinase C
Platelet Membrane Glycoproteins
Transcription Factors
Platelet Glycoprotein GPIIb-IIIa Complex
Stereoisomerism
Megakaryocytes
Membrane Glycoproteins
Electrophoretic Mobility Shift Assay
Tetradecanoylphorbol Acetate
phorbol
Thymidine
Half-Life
Proteins
Down-Regulation
Cell Proliferation

ASJC Scopus subject areas

  • Hematology
  • Cell Biology

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AP-1/c-jun and c-myc regulation during megakaryocytic differentiation of a human bi-potential growth-factor-dependent cell line. / Derigs, H. G.; Morgan, D. A.; Hoffman, R.; Litz, S. L.; Srour, Edward; Brandt, J. E.; Boswell, H.

In: Platelets, Vol. 6, No. 1, 1995, p. 24-30.

Research output: Contribution to journalArticle

Derigs, H. G. ; Morgan, D. A. ; Hoffman, R. ; Litz, S. L. ; Srour, Edward ; Brandt, J. E. ; Boswell, H. / AP-1/c-jun and c-myc regulation during megakaryocytic differentiation of a human bi-potential growth-factor-dependent cell line. In: Platelets. 1995 ; Vol. 6, No. 1. pp. 24-30.
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AU - Derigs, H. G.

AU - Morgan, D. A.

AU - Hoffman, R.

AU - Litz, S. L.

AU - Srour, Edward

AU - Brandt, J. E.

AU - Boswell, H.

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AB - Terminal megakaryocytic development is characterized by nuclear poly ploidization, appearance of specific granules, and enhanced expression of membrane platelet glycoproteins. We utilized a human GM-CSF-dependent cell line, MB-02, which is capable of under going megakaryocytic differentiation, to examine the molecular events underlying this process. The responses of MB-02 to the protein kinase C (PKC) agonist, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) were examined. GM-CSF dependent proliferation of MB-02, as measured by 3H-thymidine uptake, was greater than 95% inhibited by TPA (16 nM), but was not affected by the inactive stereoisomer, 4-αphorbol-12,13-didecanoate (4-αPDD). Transient exposure of cells to GM-CSF after growth factor deprivation led to rapid, high-level expression of normal-sized c-myc mRN A transcripts above baseline. C-myc expression was turned off by TPA (16 nM) stimulation of cells within 2-4 h. This TPA-mediated effect likely occurred at the transcriptional level since the half life of c-myc mRN A induced by GM-CSF was less than 30 min. Treatment of cells with TPA was associated with induction of c-jun and junB mRN A within 1-4 h. The protein products of these transcription factors are known to be part of the transcription factor complex Activator protein 1 (AP-1). Indeed, our data prove a rapid induction of AP-1 protein after TPA stimulation, as shown by mobility shift assays. In addition, TPA treatment resulted in expression of platelet surface glycoprotein IIb/IIIa complex (gpIIb/IIIa). These studies suggest a link between PKC stimulation by TPA and AP-1 activation with downregulation of c-myc transcription on a molecular level. At the cellular level, PKC activation was related to the acquisition of several features of the megakaryocyte development programme associated with the switch from cell proliferation to maturation.

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