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Differential induction of apoptosis in undifferentiated and differentiated
HL-60 cells by DNA topoisomerase I and II inhibitors
E Solary, R Bertrand, KW Kohn and Y Pommier
Laboratory of Molecular Pharmacology, National Cancer Institute, National
Institutes of Health, Bethesda, MD 20892.
The effects of monocytic/macrophage and granulocytic differentiation
induced by phorbol myristate acetate (TPA) and all-trans retinoic acid,
respectively, were tested on the induction of apoptosis in human
promyelocytic leukemia HL-60 cells treated with topoisomerase I and II
inhibitors. Using a filter-binding assay, we observed a strong inhibition
of DNA fragmentation induced by 3- and 24-hour continuous exposure to
camptothecin, VP-16, VM-26, and m-AMSA in TPA- differentiated cells. The
inhibition of the typical internucleosomal DNA fragmentation was confirmed
by agarose gel electrophoresis. By contrast, drug-induced DNA fragmentation
was not inhibited in retinoic acid-differentiated cells, and apoptosis
occurred in these cells after 4 to 5 days in the absence of drug treatment.
The TPA inhibitory effect was maximal after 24 hours of treatment and was
correlated with differentiation, because phorbol dibutyrate ester was
active, whereas 4- alpha-TPA, a nontumor promoter that does not induce
differentiation, was not active. Using alkaline elution, we observed that
TPA and retinoic acid differentiation were associated with changes in
topoisomerase-mediated DNA breaks that were not correlated with their
differential effects on drug-induced DNA fragmentation. Moreover, TPA also
inhibited DNA fragmentation induced by vinblastine, cycloheximide,
calphostin C, and x-rays. Using a cell-free system, we observed that DNA
fragmentation was not inhibited in nuclei from TPA-differentiated cells.
Rather, inhibition of apoptosis seemed to take place in the cytoplasm. We
conclude that phenotypic changes associated with TPA- induced
differentiation include inactivation of a cytoplasmic activity that can
induce DNA fragmentation associated with apoptosis.
Volume 81,
Issue 5,
pp. 1359-1368,
03/01/1993
Copyright © 1993 by The American Society of Hematology
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