Document Detail


Stem cell factor influences the proliferation and erythroid differentiation of the MB-02 human erythroleukemia cell line by binding to a high-affinity c-kit receptor.
MedLine Citation:
PMID:  7687159     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Stem cell factor (SCF) acts in synergy with other growth factors such as erythropoietin (Epo), granulocyte-macrophage colony-stimulating factor (GM-CSF), or interleukin-3 (IL-3), to stimulate the growth of primitive hematopoietic cells. Because of the prominent role of CSF in the maintenance of normal erythropoiesis in vivo, we examined the effects of SCF on the Epo-inducible human erythroleukemia cell line MB-02, and characterized the c-kit receptor in these cells. MB-02 cells cultured in serum-containing media do not survive in the absence of exogenous growth factors, but the addition of SCF, Epo, or IL-3 as a single factor enhanced MB-02 survival. Furthermore, in the presence of Epo, SCF (5 to 25 ng/mL) enhanced MB-02 proliferation in a dose-dependent manner, and increased the relative and absolute number of benzidine-positive cells generated. SCF also enhanced cell proliferation in the presence of either IL-3 or low concentrations of GM-CSF. A neutralizing anti-c-kit receptor monoclonal antibody (SR-1) blocked binding of 125I-SCF to MB-02 cells by 98%, and the effect of SCF on MB-02 growth, c-kit receptor-binding parameters were quantitated by equilibrium-binding experiments with 125I-SCF. MB-02 cells display a single class of high-affinity (50 pmol/L) c-kit receptors, with approximately 8,000 receptors per cell. The molecular weight of the c-kit receptor was determined by affinity cross-linking 125I-SCF to MB-02 cells. 125I-SCF-c-kit receptor complexes of approximately 155,000 and approximately 310,000 daltons were found, likely representing the monomeric and dimeric forms of the c-kit receptor. The binding affinity and molecular weight of the c-kit receptor on MB-02 cells are similar to those of normal human marrow cells. These results suggest that SCF synergizes with Epo to influence not only the proliferation but the erythroid differentiation of MB-02 cells. Thus, the MB-02 cell line may be a useful model in which to investigate the molecular mechanisms of SCF action.
Authors:
V C Broudy; D A Morgan; N Lin; K M Zsebo; F W Jacobsen; T Papayannopoulou
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Blood     Volume:  82     ISSN:  0006-4971     ISO Abbreviation:  Blood     Publication Date:  1993 Jul 
Date Detail:
Created Date:  1993-08-17     Completed Date:  1993-08-17     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7603509     Medline TA:  Blood     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  436-44     Citation Subset:  AIM; IM    
Affiliation:
Department of Medicine, University of Washington, Seattle 98195.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal
Cell Differentiation
Cell Division
Cell Survival
Erythropoietin / pharmacology
Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
Hematopoietic Cell Growth Factors / metabolism,  pharmacology*
Humans
Interleukin-3 / pharmacology
Leukemia, Erythroblastic, Acute / metabolism,  pathology*
Molecular Weight
Proto-Oncogene Proteins / chemistry,  genetics,  metabolism*
Proto-Oncogene Proteins c-kit
RNA, Messenger / metabolism
Stem Cell Factor
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
DK31232/DK/NIDDK NIH HHS; DK44194/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Hematopoietic Cell Growth Factors; 0/Interleukin-3; 0/Proto-Oncogene Proteins; 0/RNA, Messenger; 0/Stem Cell Factor; 11096-26-7/Erythropoietin; 83869-56-1/Granulocyte-Macrophage Colony-Stimulating Factor; EC 2.7.10.1/Proto-Oncogene Proteins c-kit

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