Prevention of phorbol ester receptor down modulation in human myeloblastic leukemia ML-1 cells by differentiation-stimulating serum components.

The phorbol esters 12-O-tetradecanoylphorbol-13-acetate, phorbol 12,13-didecanoate, phorbol 12,13-dibutyrate (PDB), and phorbol 12,13-dibenzoate were found to compete with [20-3H]-PDB binding to human myeloblastic leukemia ML-1 cells in approximate proportion to their differentiation-inducing capacity. Fetal bovine serum decreased the down modulation of phorbol ester receptor sites on these cells and increased PDB-induced differentiation. These two activities coeluted upon chromatography of fetal bovine serum on a Sephadex G-150 column. A partially purified fraction from pokeweed mitogen-stimulated human leukocyte-conditioned medium which effectively induced ML-1 cell differentiation also prevented the down modulation of PDB receptors. As indicated by Scatchard analysis, prevention of down modulation was due to stabilization of the number of binding sites rather than to a change in receptor affinity. In view of the previously observed modulation of growth factor binding by phorbol esters, the currently described alteration of phorbol ester receptor activity by differentiation-inducing factors implies an interaction between growth and differentiation factors in receptor modulation.