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Retinoic acid and granulocyte colony-stimulating factor synergistically
induce leukocyte alkaline phosphatase in acute promyelocytic leukemia cells
M Gianni, M Terao, S Zanotta, T Barbui, A Rambaldi and E Garattini
Unita di Biologia Molecolare, Centro Catullo e Daniela Borgomainerio,
Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy.
In this report we show a strong synergistic interaction between granulocyte
colony-stimulating factor (G-CSF) and all-trans retinoic acid (ATRA) on the
expression of leukocyte alkaline phosphatase (LAP) in freshly isolated
acute promyelocytic leukemia (APL) blasts as well as in NB40 and HL-60 cell
lines. The strong synergism observed in these cell types was not evident in
two acute leukemia cell lines (K562 and GF-D8), in normal granulocytes, and
in monocytes. In freshly isolated leukocytes derived from chronic
myelogenous leukemia (CML), in the stable phase of the disease, a weaker
interaction between ATRA and G- CSF was documented. The cross-talk between
the cytokine and the retinoid was studied in detail in NB4, an immortalized
APL leukemia cell line, retaining the 15-17 chromosomal translocation
involving the retinoic acid receptor type alpha. The treatment of NB4 cells
with G- CSF alone or ATRA alone leads to no increase and to minor induction
in LAP activity, respectively. If the cells are treated with the two
compounds simultaneously, a dramatic elevation of LAP is observed after 4
days. The synergism between G-CSF and ATRA is evident at concentrations of
the retinoid between 10(-7) and 10(-5) mol/L and at concentrations of the
cytokine between 1 and 10 ng/mL. The simultaneous presence of the two
compounds is necessary to obtain maximal increase of LAP activity and the
effect is cell density-dependent. Synergism is specific for G-CSF, and it
is not observed with other cytokines and functional inducers of the
granulocyte. The augmentation of LAP activity is the consequence of an
increased transcriptional rate of the liver/bone/kidney-type (L/B/K-type)
alkaline phosphatase gene, as determined by Northern blotting and nuclear
run-on analysis using specific cDNA probes. Only one of the two possible
alternatively spliced forms of L/B/K-type alkaline phosphatase transcript
is detected in NB4 cells after stimulation with G-CSF and ATRA. This mRNA
form, which is the one observed in normal polymorphonuclear leukocytes,
contains the most upstream leader exon. In NB4 cells, ATRA induces G- CSF,
alpha, and beta retinoic acid receptor transcripts, whereas G-CSF has minor
effects on the expression of these mRNAs.
Volume 83,
Issue 7,
pp. 1909-1921,
04/01/1994
Copyright © 1994 by The American Society of Hematology
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