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The expression, characterization and roles of macrophage colony-stimulating factor receptor in human leukemia cell
Shengsong Tang1, Guibin Chen, Qing Rao
1State Key Laboratory of Experimental Hematology, Institute Of Hematology, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin 300020, China.
Objective:
To explore the expression, characteristics and roles of macrophage colony-stimulating factor receptor (M-CSF-R) in human leukemia cell lines.
Methods:
Peripheral blood mononuclear cells (PBMCs) collected from 3 healthy persons, cord blood mononuclear cells (CBMCs) collected from 5 healthy persons and 4 human myelomonocytic leukemia cell lines including J6-1, J6-2, K562 and HL-60 were studied by using ABC immunoperoxidaes assay, indirect immunofluorescene staining, flow cytometry, and Western blot.
Results:
M-CSF-R was noticed to be localized in the cytoplasm, nucleus and at the membrane in 4 human leukemia cell lines; expression of M-CSF-R was not detected in normal human PBMCs without PHA stimulation. Human PBMCs stimulated by PHA expressed a low level of M-CSF-R. Frequencies of membrane bound M-CSF-R (M-CSF-mR) expression in J6-1, J6-2, K562 and HL-60 were 78.9%, 72.6%, 54.9% and 58.0% respectively. Frequencies of cytoplasm and nucleus associated M-CSF-R (M-CSF-cnR) were 52.3%, 44.3%, 28.0% and 65.3% respectively. One form of M-CSF-R with a molecular weight of 120 000 was detected both in the cytoplasm and nucleus of HL-60 cells. The half-life of M-CSF-cnR in leukemia cells mentioned above was longer than that of corresponding M-CSF-R in stimulated CBMCs, and the half-life of M-CSF-mR in leukemia cells was extended except that of M-CSF-mR in K562 cells. Both anti-M-CSF-R monoclonal antibody and recombinant human M-CSF soluble receptor could cause the growth arrest of HL-60 cell in G(0)/G(1) phase, and could inhibit the formation of colony of HL-60 cell in soft agarose.
Conclusions:
Expression of M-CSF-R in leukemia cells is heterogeneous. The accumulation of cellular M-CSF-R results in the low degradation rate of cellular M-CSF-R in leukemia cells, which could be a potential mitotic signal. Signal mediated by M-CSF-R is important and necessary for the growth of HL-60 cell.