Difference in response to colony-stimulating factors and involvement of protein kinase C signal transduction system in three subclones from the ME-1 cell line and two sublines.
We previously established a cell line from a patient with acute myelomonocytic leukemia with eosinophilia (M4E0), ME-1. ME-1 cells are responsive to colony-stimulating factors (CSFs) such as interleukin-3 (IL-3), IL-4, and granulocyte-macrophage CSF (GM-CSF), and exhibit monocyte-macrophage differentiation. We isolated three subclones, ME-F1 from ME-1, and ME-F2 and ME-F3 from two sublines of ME-1. These subclones had different morphologic, cytochemical, phenotypic, and cytogenetic features. They represented different monocytic-lineage differentiation stages and exhibited different responses to IL-3, GM-CSF, and especially IL-4. IL-3, GM-CSF, and IL-4 enhanced proliferation and differentiation to macrophage-like cells in the ME-F1 subclone. However, they enhanced only proliferation of ME-F2 cells and only differentiation to macrophage-like cells in the ME-F3 subclone. To elucidate possible differences in signal transduction mechanisms in ME-F1, ME-F2, and ME-F3 cells following stimulation by CSFs, we studied the effects of IL-3 and IL-4 on protein kinase C (PKC) activity. Both IL-3 and IL-4 induced a rapid, transient decrease of cytosolic PKC in ME-F1 cells, but did not affect PKC activity in ME-F2 and ME-F3 cells. The PKC inhibitors, 1-(5-isoquinolinyl-sulfonyl)-2-methylpiperazine (H-7) and calphostin C inhibited IL-3-induced enhancement of proliferation and differentiation of ME-F1 cells, but did not inhibit enhancement of proliferation of ME-F2 cells and differentiation of ME-F3 cells. Our data suggest that PKC-dependent signal transduction is considerably related to IL-3-induced proliferation and differentiation of ME-F1 cells. In addition, it was demonstrated that the two subclones, ME-F2 and ME-F3, lost one of the two responses of ME-F1 cells to CSFs, either proliferation or differentiation, and simultaneously lost PKC-dependent response to CSFs.
First Department of Internal Medicine, School of Medicine, Ehime University, Japan.
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K
Yanagisawa
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M
Sato
T
T
Horiuchi
H
H
Hasegawa
S
S
Fujita
D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002869:Chromosome Aberrations; D003115:Colony-Stimulating Factors / Q000494:pharmacology*; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D015479:Leukemia, Myelomonocytic, Acute / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D011493:Protein Kinase C / Q000502:physiology*; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
D003115:Colony-Stimulating Factors; D007377:Interleukin-3; D015847:Interleukin-4; D011493:Protein Kinase C