All trans retinoic acid induces apoptosis in acute promyelocytic NB4 cellswhen combined with isoquinolinediol, a poly(ADP-ribose) polymerase inhibitor

NB4 cells, a model of acute promyelocytic leukemia have been shown to under go granulocytic differentiation in response to all trans retinoic acid (ATR A), or monocytic differentiation in response to 1 alpha,25 dihydroxyvitamin D-3 (1,25 D-3) and phorbol ester. We have shown previously that the expres sion of poly(ADP-ribose) polymerase (PARP) is dramatically increased during monocytic differentiation and completely down-regulated during neutrophili c differentiation. Here we show that NB4 cells simultaneously treated with ATRA and isoquinolinediol (Iso-Q), a specific PARP inhibitor, fail to diffe rentiate into neutrophils. Nitroblue tetrazolium (NBT) dye reduction was in hibited by 48% and neutrophil formation was reduced by 75%. NB4 cells treat ed with ATRA and Iso-Q instead showed features of apoptosis including nucle ar condensation, DNA fragmentation, and PARP cleavage yielding a 85 kDa fra gment. NB4 cells treated with ATRA in combination with 1,25 D-3, a monocyti c differentiation inducer, while continuing to reduce NET also failed to ma ture into neutrophils or monocytes and again showed features of apoptosis. Down-regulation of Bcl-2 protein expression was evident in NB4 cells treate d with ATRA and ATRA in combination with Iso-Q or 1,25 D-3, but not in cell s treated with a classic chemotherapeutic agent, arabinosycytosine, suggest ing that Bcl-2 down-regulation is neither necessary, nor specific for apopt osis in this model. (C) 2000 Elsevier Science Ltd. All rights reserved.