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Cytokine signals through STAT3 promote expression of granulocyte secondary granule proteins in 32D cells.

Publication ,  Journal Article
Wang, L; Arcasoy, MO; Watowich, SS; Forget, BG
Published in: Exp Hematol
March 2005

OBJECTIVE: In a previous study, we showed that activation of a transfected human erythropoietin receptor (EPOR) in the murine myeloid cell line 32D resulted in the development of morphologic features of granulocytic differentiation and expression of the neutrophil primary granule protein myeloperoxidase. We now studied if EPOR signaling could also mediate secondary granule protein gene expression and investigated the signal transduction requirements for induction of secondary granule gene expression in 32D cells. MATERIALS AND METHODS: Wild-type and variant 32D cells expressing normal or chimeric EPORs or receptors for granulocyte colony-stimulating factor (G-CSFRs) were stimulated with EPO or G-CSF and the expression of granulocyte-specific genes was analyzed by Northern blot analysis. To determine the signaling mechanisms required for secondary granule protein gene induction, the activation of STAT pathways following growth factor stimulation was studied by Western blot analysis. RESULTS: We found that EPO treatment of 32D cells engineered to express EPOR did not result in induction of the secondary granule protein genes encoding lactoferrin and 24p3 lipocalin, the mouse homolog of human N-Gal, or the myeloid transcription factor C/EBPepsilon. Replacement of the intracellular domain of EPOR with the intracellular domain of G-CSFR in a chimeric receptor was associated with EPO-mediated induction of lactoferrin, 24p3 lipocalin, and C/EBPepsilon genes. We found that STAT3 phosphorylation was mediated by the intracellular domain of G-CSFR, but not EPOR. Replacement of one or two of the STAT5 binding sites in the intracytoplasmic domain of the EPOR with STAT3 binding sites resulted in EPO-mediated STAT3 activation and a marked increase in the expression of the 24p3 lipocalin gene. Knockdown of STAT3 protein levels with siRNA caused significant decrease in 24p3 lipocalin gene induction. CONCLUSION: These results indicate that EPOR signaling cannot substitute for G-CSFR signaling to stimulate secondary granule protein gene expression in 32D cells. In addition, STAT3 is a critical mediator of 24p3 lipocalin gene expression in these cells.

Duke Scholars

Published In

Exp Hematol

DOI

ISSN

0301-472X

Publication Date

March 2005

Volume

33

Issue

3

Start / End Page

308 / 317

Location

Netherlands

Related Subject Headings

  • Transcriptional Activation
  • Trans-Activators
  • Signal Transduction
  • STAT3 Transcription Factor
  • Recombinant Proteins
  • Receptors, Granulocyte Colony-Stimulating Factor
  • Receptors, Erythropoietin
  • Protein Biosynthesis
  • Mice
  • Immunology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Wang, L., Arcasoy, M. O., Watowich, S. S., & Forget, B. G. (2005). Cytokine signals through STAT3 promote expression of granulocyte secondary granule proteins in 32D cells. Exp Hematol, 33(3), 308–317. https://doi.org/10.1016/j.exphem.2004.11.014
Wang, Lei, Murat O. Arcasoy, Stephanie S. Watowich, and Bernard G. Forget. “Cytokine signals through STAT3 promote expression of granulocyte secondary granule proteins in 32D cells.Exp Hematol 33, no. 3 (March 2005): 308–17. https://doi.org/10.1016/j.exphem.2004.11.014.
Wang L, Arcasoy MO, Watowich SS, Forget BG. Cytokine signals through STAT3 promote expression of granulocyte secondary granule proteins in 32D cells. Exp Hematol. 2005 Mar;33(3):308–17.
Wang, Lei, et al. “Cytokine signals through STAT3 promote expression of granulocyte secondary granule proteins in 32D cells.Exp Hematol, vol. 33, no. 3, Mar. 2005, pp. 308–17. Pubmed, doi:10.1016/j.exphem.2004.11.014.
Wang L, Arcasoy MO, Watowich SS, Forget BG. Cytokine signals through STAT3 promote expression of granulocyte secondary granule proteins in 32D cells. Exp Hematol. 2005 Mar;33(3):308–317.
Journal cover image

Published In

Exp Hematol

DOI

ISSN

0301-472X

Publication Date

March 2005

Volume

33

Issue

3

Start / End Page

308 / 317

Location

Netherlands

Related Subject Headings

  • Transcriptional Activation
  • Trans-Activators
  • Signal Transduction
  • STAT3 Transcription Factor
  • Recombinant Proteins
  • Receptors, Granulocyte Colony-Stimulating Factor
  • Receptors, Erythropoietin
  • Protein Biosynthesis
  • Mice
  • Immunology