Document Detail

Erythropoietin-specific cell cycle progression in erythroid subclones of the interleukin-3-dependent cell line 32D.
MedLine Citation:
PMID:  7679009     Owner:  NLM     Status:  MEDLINE    
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)
Y Shimada; G Migliaccio; H Ralph; A R Migliaccio; H Shaw
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Blood     Volume:  81     ISSN:  0006-4971     ISO Abbreviation:  Blood     Publication Date:  1993 Feb 
Date Detail:
Created Date:  1993-03-11     Completed Date:  1993-03-11     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7603509     Medline TA:  Blood     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  935-41     Citation Subset:  AIM; IM    
Laboratory of Hematopoietic Growth Factors, Lindsley F. Kimball Research Institute, New York Blood Center, New York 10021.
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MeSH Terms
Calcium-Binding Proteins / genetics
Cell Cycle / genetics,  physiology*
Cell Differentiation
Cell Line
Clone Cells / cytology
DNA / metabolism
Erythrocytes / cytology*,  metabolism
Erythropoietin / pharmacology*
Gene Expression
Genes, myc
Granulocyte Colony-Stimulating Factor / pharmacology
Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
Histones / genetics
Interleukin-3 / pharmacology*
Protein Kinases / metabolism
RNA, Messenger / metabolism
Grant Support
Reg. No./Substance:
0/Calcium-Binding Proteins; 0/Histones; 0/Interleukin-3; 0/RNA, Messenger; 11096-26-7/Erythropoietin; 143011-72-7/Granulocyte Colony-Stimulating Factor; 83869-56-1/Granulocyte-Macrophage Colony-Stimulating Factor; 9007-49-2/DNA; EC 2.7.-/Protein Kinases
Erratum In:
Blood 1993 Jun 1;81(11):3168
Note: Shaw H [corrected to Ralph H]

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