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Erythropoietin-specific cell cycle progression in erythroid subclones of
the interleukin-3-dependent cell line 32D [published erratum appears in
Blood 1993 Jun 1;81(11):3168]
Y Shimada, G Migliaccio, H Ralph, AR Migliaccio and H] Shaw H$[corrected to Ralph
Laboratory of Hematopoietic Growth Factors, Lindsley F. Kimball Research
Institute, New York Blood Center, New York 10021.
Recently, a variety of growth factor-dependent subclones of the murine
interleukin-3 (IL-3)-dependent cell line 32D have been isolated. These
subclones include those dependent for growth on erythropoietin (Epo) (32D
Epo), granulocyte-macrophage colony-stimulating factor (GM-CSF) (32D GM),
or granulocyte colony-stimulating factor (G-CSF) (32D G). 32D Epo1.1 is a
revertant of 32D Epo and is capable of growing in IL-3. These cell lines
express the differentiation program appropriate to the specific growth
factor and depend on the growth factors not only for proliferation but also
for survival. To determine how the signal for proliferation is triggered by
various growth factors, we examined the DNA histograms and the expression
of cell cycle-specific genes in the different cell lines. The cell
cycle-specific genes analyzed were myc (early G1), myb (late G1), and the
structural genes for the calcium- binding protein 2A9 (middle G1) and
histone H3 (G1-S boundary). The DNA histogram analysis of cells in the
logarithmic phase of growth showed that approximately 40% of 32D, 32D GM,
32D G, and 32D Epo1.1 (growing in IL-3) were cells with a 2N DNA content
(and therefore in G0/G1), and another 40% have a DNA content intermediate
between 2N and 4N (in S phase). In contrast, 32D Epo and 32D Epo1.1
(growing in Epo) had fewer cells in the G0/G1 phase of the cell cycle
compared with the number of cells that were in the S phase (19% to 31% v
69% to 78%, respectively). Because all the cell lines have comparable
doubling times (15 to 18 hours), the cell distribution among the phases of
the cell cycle is proportional to the length of the phase. Therefore, cells
growing in IL- 3 (32D and 32D Epo1.1), GM-CSF (32D GM), or G-CSF (32D G)
progress along the cycle in a manner typical of previously reported
nontransformed cell lines. In contrast, cells growing in Epo (32D Epo or
32D Epo1.1) spend relatively less time in G0/G1 and correspondingly more
time in S. These data were confirmed by the analysis of the tritiated
thymidine (3H-TdR) suicide rate and of the expression of cell
cycle-specific genes. The 32D and 32D Epo1.1 cells growing in IL-3 had a
suicide rate of congruent to 50%, whereas the suicide rate of 32D Epo and
32D Epo1.1 growing in Epo was higher than 75%.(ABSTRACT TRUNCATED AT 400
WORDS)
Volume 81,
Issue 4,
pp. 935-941,
02/15/1993
Copyright © 1993 by The American Society of Hematology
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