title: 1,25-Dihydroxyvitamin D3 and phorbol myristate acetate produce divergent phenotypes in a monomyelocytic cell line.
abstract: The human monomyelocytic HL-60 cell line differentiates along a monocytic lineage when cultured in the presence of phorbol myristate acetate (PMA) or 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. The protooncogene c-fms, coding for the macrophage colony-stimulating factor (M-CSF) receptor, is characteristically expressed in cells of monocytic lineage, but is not present in HL-60 cells. Since M-CSF has an undefined role in osteoclast development, we wished to examine the effect of 1,25-(OH)2D3, a strong promoter of osteoclast development, on the induction of M-CSF receptor. Treatment of HL-60 cells with 10-50 ng/ml PMA for 3 days stimulated expression of c-fms, as measured by steady state mRNA levels. Treatment with 10 nM 1,25-(OH)2D3 for 4-72 h did not promote c-fms expression. In fact, 1,25-(OH)2D3 attenuated PMA-stimulated c-fms expression in a dose-dependent manner (ED50, 1 nM). Along with attenuation of c-fms expression, 1,25-(OH)2D3 induced expression of mRNA for the osteoclastic enzyme carbonic anhydrase-II (CA II) almost 2-fold over the level expressed in untreated HL-60 cells. The addition of PMA to the culture diminished the basal expression of CA II mRNA, but did not affect 1,25-(OH)2D3-stimulated CA II induction. PMA and 1,25-(OH)2D3, thus, promote divergent phenotype development in the HL-60 cell. Induction of c-fms mRNA by PMA should ensure further macrophage development. The effect of 1,25-(OH)2D3 to attenuate expression of c-fms mRNA and stimulate CA II mRNA suggests that 1,25-(OH)2D3 activates osteoclast, rather than macrophage, development. 1,25-(OH)2D3-induced phenotypic changes may, therefore, involve modulation of the M-CSF effect.
authors.affiliation: Department of Medicine, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: D M
authors.initials: DM
authors.lastname: Biskobing
authors.forename: J
authors.initials: J
authors.lastname: Rubin
mesh_terms: D001483:Base Sequence; D002117:Calcitriol / Q000494:pharmacology*; D002256:Carbonic Anhydrases / Q000096:biosynthesis* / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects*; D015342:DNA Probes; D004305:Dose-Response Relationship, Drug; D016236:Genes, fms / Q000187:drug effects*; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D015345:Oligonucleotide Probes; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000187:drug effects / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D015342:DNA Probes; D007527:Isoenzymes; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D016186:Receptor, Macrophage Colony-Stimulating Factor; D002256:Carbonic Anhydrases; D002117:Calcitriol; D013755:Tetradecanoylphorbol Acetate