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c-kit expression by CD34+ bone marrow progenitors and inhibition of
response to recombinant human interleukin-3 following exposure to c-kit
antisense oligonucleotides
JP Catlett, JA Leftwich, EH Westin, S Grant and TF Huff
Department of Hematology/Oncology, Medical College of Virginia, Richmond
23298.
The c-kit proto-oncogene encodes a receptor having tyrosine-specific kinase
activity and has been mapped to chromosome 4 in the human and chromosome 5
in the mouse, at the dominant white spotting locus (W). Mutations at the W
locus affect various aspects of murine hematopoiesis. The c-kit
proto-oncogene has been shown to be expressed by leukemic myeloblasts, but
not by normal unseparated human bone marrow cells. The role of this
oncogene in differentiation and proliferation of human hematopoietic
progenitors is presently undefined. To determine c-kit expression by normal
hematopoietic progenitors, CD34+ cells were isolated from disease-free
human bone marrow, and RNA-based polymerase chain reaction (PCR) techniques
were used to assess expression. By this method, we have demonstrated c-kit
expression by CD34+ bone marrow progenitors. To address the functional
requirement for c-kit expression in normal human hematopoiesis, CD34+ cells
were incubated in the presence of sense, antisense, or missense
oligonucleotides to c-kit, and subsequently cultured in the presence of
either recombinant human granulocyte-macrophage colony-stimulating factor
(rhGM-CSF) or recombinant human interleukin-3 (rhIL-3). Exposure of CD34+
cells to c-kit antisense oligonucleotides significantly inhibited
colony-forming ability of cells cultured in the presence of rhIL-3, but had
no effect on colony formation of cells cultured in rhGM- CSF. Together,
these data suggest a possible role for c-kit in hematopoietic proliferation
and differentiation that may be linked to some, but not all, stimulatory
factors.
Volume 78,
Issue 12,
pp. 3186-3191,
12/15/1991
Copyright © 1991 by The American Society of Hematology
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