title: Effects of bryostatin 1 and calcium ionophore (A23187) on apoptosis and differentiation in human myeloid leukemia cells (HL-60) following 1-beta-D-arabinofuranosylcytosine exposure.
abstract: The goal of the present study was to determine whether partial restoration of the differentiation-inducing capacity of the PKC activator bryostatin 1 by the calcium ionophore A23187 is accompanied by enhancement of apoptosis in ara-C-pretreated human leukemia cells. When HL-60 cells were exposed to ara-C (10 or 100 microM;6 h) followed by bryostatin 1 alone (10 nM; 24 h), no increase in apoptosis was noted. In contrast, subsequent exposure of ara-C-pretreated cells to A23187 (250 nM; 24 h) increased apoptosis by approximately 100%. When ara-C-pretreated cells were incubated with A23187 and bryostatin 1, no further potentiation of cell death (compared to cells exposed to A23187 alone) was observed. Nevertheless, the combination of bryostatin 1 and A23187 substantially increased inhibition of clonogenicity in cells preincubated with ara-C (e.g., by > or = 2 logs). This effect was associated with morphological and functional evidence (i.e., plastic adherence) of enhanced leukemic cell maturation. The differentiating capacity of the combination of bryostatin 1 and A23187 was significantly weaker than that of the phorbol diester, PMA (10 nM), and unaccompanied (at 24 h) by induction of the cyclin-dependent kinase inhibitors (CDKIs) p21WAF1/CIP1 and p27KIP1. However, the extent of apoptosis was comparable in cells exposed to ara-C followed by PMA or bryostatin 1 + A23187, suggesting that differentiation per se is not solely responsible for enhancement of cell death in ara-C-pretreated cells. Coadministration of bryostatin 1 and the organotellurium compound AS101, which mimics the actions of A23187 in some systems, after ara-C also led to enhanced antiproliferative effects which were unaccompanied by an increase in apoptosis. Finally, exposure of cells to ara-C followed by other differentiation-inducing agents, including dimethylsulfoxide and sodium butyrate also resulted in increases in cell death in this cell line. These findings indicate that the inability of bryostatin 1 to potentiate apoptosis in ara-C-pretreated HL-60 cells may involve factors other than an inadequate differentiation stimulus. They also suggest that loss of leukemic self-renewal capacity following exposure to cytotoxic and differentiation-inducing agents may involve mechanisms other than, or in addition to, potentiation of apoptosis, particularly cellular maturation.
authors.affiliation: Department of Medicine, Medical College of Virginia, Richmond VA, 23298, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Vrana
authors.forename: A S
authors.initials: AS
authors.lastname: Rao
authors.forename: Z
authors.initials: Z
authors.lastname: Wang
authors.forename: W D
authors.initials: WD
authors.lastname: Jarvis
authors.forename: S
authors.initials: S
authors.lastname: Grant
mesh_terms: D000081:Acetamides / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D054713:Bryostatins; D000001:Calcimycin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D003561:Cytarabine / Q000494:pharmacology*; D053938:DNA Fragmentation; D018922:HL-60 Cells; D006801:Humans; D007783:Lactones / Q000494:pharmacology*; D018942:Macrolides; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000081:Acetamides; D000970:Antineoplastic Agents; D054713:Bryostatins; D007783:Lactones; D018942:Macrolides; D003561:Cytarabine; D000001:Calcimycin; C046785:bryostatin 1; C014026:hexamethylene bisacetamide; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium