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Endocrinology, Vol 132, 862-866, Copyright © 1993 by Endocrine Society

ARTICLES

1,25-Dihydroxyvitamin D3 and phorbol myristate acetate produce divergent phenotypes in a monomyelocytic cell line

DM Biskobing and J Rubin
Department of Medicine, Emory University School of Medicine, Atlanta, Georgia 30322.

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.


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