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J Biol Chem, Vol. 274, Issue 36, 25471-25480, September 3, 1999

Commitment of Neutrophilic Differentiation and Proliferation of HL-60 Cells Coincides with Expression of Transferrin Receptor
EFFECT OF GRANULOCYTE COLONY STIMULATING FACTOR ON DIFFERENTIATION AND PROLIFERATION

Toshie Kanayasu-Toyoda, Teruhide Yamaguchi, Eriko Uchida, and Takao Hayakawa

From the Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, 158-8501 Tokyo, Japan

To examine the regulatory mechanisms of proliferation and maturation in neutrophilic lineage cells, we have tried to sort dimethyl sulfoxide (Me2SO)-treated HL-60 cells into transferrin receptor (Trf-R) positive (Trf-R+) and negative (Trf-R-) cells. Differentiated Trf-R- cells expressed more formyl-Met-Leu-Phe receptor (fMLP-receptor) and ability of Obardot 2 genaration, as markers of differentiation, than Trf-R+ cells, and Trf-R- cell differentiation was markedly accelerated by the incubation with granulocyte colony stimulating factor (G-CSF). On the other hand, Trf-R+ cells had a tendency to proliferate rather than differentiate, and proliferation was enhanced by G-CSF. These results indicate that Trf-R expression coincides with the commitment to proliferate or differentiate of HL-60 cells, and G-CSF accelerates these commitments. G-CSF-induced tyrosine phosphorylation of STAT 3 in Trf-R- cells much more than in Trf-R+ cells. Protein 70 S6 kinase expression was higher in Trf-R+ cells than in Trf-R- cells. Furthermore, p70 S6 kinase was hyperphosphorylated by G-CSF in Trf-R+ cells, but not in Trf-R- cells. Rapamycin, an inhibitor of p70 S6 kinase activity, inhibited G-CSF-dependent proliferation of Trf-R+ cells and increased fMLP-R expression on these cells. These results suggest that commitment to proliferation and differentiation in Me2SO-treated HL-60 cells is preprogrammed and correlated with Trf-R expression, and G-CSF potentiates the cellular commitment. STAT 3 may promote differentiation of Me2SO-treated HL-60 cells into neutrophils, while p70 S6 kinase may promote proliferation and negatively regulate neutrophilic differentiation.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.


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