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Blood, Vol. 94 No. 2 (July 15), 1999:
pp. 560-571
C/EBP Bypasses Granulocyte Colony-Stimulating Factor Signals to
Rapidly Induce PU.1 Gene Expression, Stimulate Granulocytic
Differentiation, and Limit Proliferation in 32D cl3 Myeloblasts
Xinping Wang,
Edward Scott,
Charles L. Sawyers, and
Alan D. Friedman
From the Johns Hopkins Oncology Center, Division of Pediatric
Oncology, Baltimore, MD; the Institute for Human Gene Therapy,
University of Pennsylvania, Philadelphia, PA; and the Molecular Biology
Institute and Department of Medicine, University of California, Los
Angeles, CA.
Within hematopoiesis, C/EBP is expressed only in myeloid cells,
and PU.1 is expressed mainly in myeloid and B-lymphoid cells. C/EBP-deficient mice lack the neutrophil lineage and retain
monocytes, whereas PU.1-deficient mice lack monocytes and have severely
reduced neutrophils. We expressed a C/EBP-estrogen receptor
ligand-binding domain fusion protein, C/EBPWT-ER, in 32D cl3
myeloblasts. 32D cl3 cells proliferate in interleukin-3 (IL-3) and
differentiate to neutrophils in granulocyte colony-stimulating factor
(G-CSF). In the presence of estradiol, C/EBPWT-ER induced
morphologic differentiation and the expression of the myeloperoxidase,
lactoferrin, and G-CSF receptor mRNAs. C/EBPWT-ER also induced a
G1/S cell cycle block, with induction of p27 and Rb
hypophosphorylation. bcr-ablp210 prevented 32D cl3 cell
differentiation. Activation of C/EBP-ER in
32D-bcr-ablp210 or Ba/F3 B-lymphoid cells induced cell
cycle arrest independent of terminal differentiation. C/EBPWT-ER
induced endogenous PU.1 mRNA within 8 hours in both 32D cl3 and Ba/F3
cells, even in the presence of cycloheximide, indicating that C/EBP
directly activates the PU.1 gene. However, activation of a PU.1-ER
fusion protein in 32D cl3 cells induced myeloperoxidase (MPO) RNA but
not terminal differentiation. Thus, C/EBP acts downstream of G-CSF
and upstream of PU.1, p27, and potentially other factors to induce
myeloblasts to undergo granulocytic differentiation and cell cycle arrest.
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S. A. Burel, N. Harakawa, L. Zhou, T. Pabst, D. G. Tenen, and D.-E. Zhang
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[Abstract]
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H. Nakajima and J. N. Ihle
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Blood,
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[Abstract]
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N. Shiroshita, M. Musashi, K. Sakurada, K. Kimura, Y. Tsuda, S. Ota, H. Iwasaki, T. Miyazaki, T. Kato, H. Miyazaki, et al.
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[Abstract]
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K. R. Calvo, D. B. Sykes, M. Pasillas, and M. P. Kamps
Hoxa9 Immortalizes a Granulocyte-Macrophage Colony-Stimulating Factor-Dependent Promyelocyte Capable of Biphenotypic Differentiation to Neutrophils or Macrophages, Independent of Enforced Meis Expression
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[Abstract]
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R. Lu and P. M. Pitha
Monocyte Differentiation to Macrophage Requires Interferon Regulatory Factor 7
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[Abstract]
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