CONTROL OF MACROPHAGE CELL-DIFFERENTIATION IN HUMAN PROMYELOCYTIC HL-60 LEUKEMIA-CELLS BY 1,25-DIHYDROXYVITAMIN-D3 AND PHORBOL-12-MYRISTATE-13-ACETATE

  • 1 January 1983
    • journal article
    • research article
    • Vol. 43  (10) , 4989-4996
Abstract
Human promyelocytic leukemia cells (HL-60) were induced to differentiate into macrophage-like cells in a dose (3 .times. 10-10 to 10-7 M) and time (1-6 days)-dependent manner by 1,25-dihydroxyvitamin D3 and the tumor promoter, phorbol-12-myristate-13-acetate. Differentiation was determined by an increase in the percentage of morphologically mature cells, in lysozyme and nonspecific esterase activities, and in reactivity with the murine OKM1 monoclonal antibody. Two HL-60 cell variants, designated as R-80 and B-11, were also examined. R-80 cells, which are resistant to induction of cell differentiation by phorbol-12-myristate-13-acetate, also exhibited resistance, although to a lesser degree, to induction of cell differentiation by 1,25-dihydroxyvitamin D3. The resistance to the action of the 2 compounds is presumably not due to similar binding sites for the 2 inducers, since 1,25-dihydroxyvitamin D3 was unable to compete for the phorbol diester binding sites as measured by [3H]phorbol-12,13-dibutyrate binding. B-11 cells were resistant to induction of cell differentiation by 1,25-dihydroxyvitamin D3,phoirbol-12-myristate-13-acetate, retinoic acid, and dimethyl sulfoxide. Two-dimensional electrophoretic analysis of HL-60 cell protein patterns indicated that treatment of the HL-60 cells with 1,25-dihydroxyvitamin D3, phorbol-12-myristate-13-acetate, retinoic acid, and dimethyl sulfoxide caused the cells to express various monocyte-macrophage and granulocyte marker proteins. None of the inducers caused a protein pattern identical to that of peripheral monocytes or granulocytes in the HL-60 cells, but the protein pattern of the HL-60 cells treated with 1,25-dihydroxyvitamin D3 was the closest to that of peripheral blood monocytes. 1,25-dihydroxyvitamin D3 induces in the HL-60 cells a phenotype that resembles, but is not identical to, that of peripheral monocytes-macrophages.