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

Abstract

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.

Array CGH of fusion gene–positive leukemia‐derived cell lines reveals cryptic regions of genomic gain and loss

“…Three subclones of the ME‐1 cell line showing different morphological, cytochemical, phenotypic, and cytogenetic features have been described (Yanagisawa et al, 1991, 1994). The present study yielded a karyotype closest to that of the ME‐F2 subclone, but it also identified new regions of loss and gain not in the reported karyotype (Table 1).…”

“…These multiple rearrangements involving chromosome 11 resulted in gain of most of the long arm, 11q13.2–qter. The del(17p) in the original karyotype (Yanagisawa et al, 1994) was shown to result from an unbalanced t(2;17), giving rise to gain of 2p25.1–pter. Also found was gain of the entire long arm of chromosome 18, which resulted from an unbalanced t(9;18).…”