1,25-Dihydroxyvitamin D3 (1,25-(OH)2D3) induces monocytic differentiation of the human promyelocytic leukemia line, HL-60, and enhances Ca2+ transport in target cells of the mineral metabolism system. Hence, we determined whether the steroid's maturational effect on HL-60 involves alterations of intracellular calcium ([Ca2+](i). We found that, as detected by indo-1 fluorescence, [Ca2+](i) increases in a slow tonic manner from 99 ± 11 nM in virgin HL-60 to 182 ± 19 nM (p<0.001) in those treated with 1,25-(OH)2D3 for 24 h. The first apparent rise in [Ca2+](i) occurs at between 6 and 12 h and parallels expression of α-thrombin and N-formyl-methionyl-leucyl-phenylalanine (fMLP) receptors. This increase in [Ca2+](i) is derived from extracellular calcium as its reduction abolishes the effect. The increase in [Ca2+](i) is associated with an increase in inositol trisphosphate-stimulated Ca2+ flux from intracellular stores. Interestingly, 1,25-(OH)2D3-mediated HL-60 differentiation as manifest by expression of the macrophage-specific antigen, 63D3, is not blocked by low extracellular calcium. In contrast, the fMLP-induced superoxide ion generation is diminished if the increase in [Ca2+](i) is prevented. Furthermore, fMLP-stimulated signal transduction is also reduced by limiting the stimulation of [Ca2+](i) during 1,25-(OH)2D3 treatment. Thus, although differentiation of HL-60 to the monocytic phenotype by 1,25-(OH)2D3 is Ca2+-independent, expression of response to regulatory stimuli requires priming of cellular Ca2+ stores. The latter appears to be induced by 1,25-(OH)2D3 via stimulated Ca2+ entry through the plasma membrane.

Original languageEnglish
Pages (from-to)16039-16044
Number of pages6
JournalJournal of Biological Chemistry
Issue number31
StatePublished - 1988


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