Activation and growth of colony-stimulating factor-dependent cell lines is cell cycle stage dependent.

Abstract

Hematopoietic cell development is regulated by a series of growth factors that are progressively restricted in their biological activity. IL-3 is a multi-lineage growth factor that supports the growth and differentiation of progenitor cells belonging to multiple lineages. However, the mechanism by which IL-3 induces proliferation and differentiation of these cells is not completely understood. In this report, we have used two IL-3-dependent cell lines, FDC-P1 (a myeloid progenitor) and F15.12 (a lymphoid progenitor) to investigate IL-3-mediated growth and differentiation. When either FDC-P1 or FL5.12 cells are deprived of IL-3, greater than 90% of all cells accumulate in the G0 phase of the cell cycle. Upon readdition of IL-3, the cells will reenter the active phases of the cell cycle. Therefore, IL-3 can act as both a competence (G0----G1) factor, and a progression (G1----M) factor for hematopoietic precursor clones. FDC-P1 cells can also proliferate in response to granulocyte/macrophage colony-stimulating factor (G/M-CSF) and IL-4 (B cell stimulatory factor 1 [BSF-1]). However, resting (G0) FDC-P1 cells have lost their ability to grow in response to both G/M-CSF and IL-4, even though both factors can induce a G0----G1 transition. Therefore, G/M-CSF or IL-4 behave as progression factors among certain IL-3-responsive clones, and in those cases only in defined points in the cell cycle. Both IL-4 and G/M-CSF can maintain long-term growth of FDC-P1 cells. Upon removal of factor for 24 h, these clones accumulate in the G1 phase of the cell cycle and do not appear to enter G0 even after 36 h of factor deprivation. Therefore, cells maintained in G/M-CSF or IL-4 have altered growth requirements compared with the IL-3-dependent lines from which they were derived. The ability of various hematopoietic growth factors to regulate cell cycle progression in IL-3-dependent cell lines is dependent not only upon the lineage from which these cells were derived, but also the phase of the cell cycle in which those cells reside. The consequences of these interactions dictate the manner by which various clones will respond to CSFs and whether the cells will grow and/or differentiate.

Interactions of dimethyl sulfoxide and granulocyte‐macrophage colony‐stimulating factor on the cell cycle kinetics and phosphoproteins of G1‐enriched HL‐60 cells: Evidence of early effects on lamin B phosphorylation

“…Our finding, suggesting that GM-CSF stimulates GI to S-Dhase Droaession of HL-60 cells. Drovides the first data-on the egects of this cytokine 0; a synchronized human cell-line and extends results obtained with IL-3lGM-CSF-dependent mouse cell-lines and CSF-1dependent mouse peritoneal macrophages, where these hematokines also have been found to promote G, to S progression (Pluznik et al, 1984;Kelvin et al, 1986;London and McKearn, 1987;Hasthorpe et al, 1988;Tushinski and Stanley, 1985). We found that GM-CSF addition could be delayed for at least half of G, without compromising progression in the next cell cycle.…”

Proteomic and Metabolomic Characterization of a Mammalian Cellular Transition from Quiescence to Proliferation

“…There is also a decrease in RNA content as measured by acridine orange (AO) staining, another reported characteristic of the G0 state (Fig. 1C) (London and McKearn, 1987). …”

Inhibition of apoptosis by the expression of antisense Nedd2

“…Murine myeloid progenitor cell line FDC-P1, which also expressed moderate levels of Nedd2 mRNA (Fig. 2) was chosen for the antisense studies using mouse Nedd2 cDNA expression constructs, because of its apoptosis response has been well characterized [29,30]. This cell line can be maintained with either GM-CSF or IL-3 and upon removal of the factor undergoes apoptosis within 1 2 days [29,30].…”

“…2) was chosen for the antisense studies using mouse Nedd2 cDNA expression constructs, because of its apoptosis response has been well characterized [29,30]. This cell line can be maintained with either GM-CSF or IL-3 and upon removal of the factor undergoes apoptosis within 1 2 days [29,30]. FDC-P1 cells were transfected by lipofection with a mammalian expression vector (pCXN2), the vector carrying Nedd2 cDNA containing entire coding region in sense orientation (PCXN2-N2), or 5'-1.0 kb fragment of the Nedd2 cDNA in antisense orientation (pCXN2-N2AS).…”