Continuous suppression of globin gene expression and differentiation of Friend erythroleukemia cells by phorbol 12-myristate 13-acetate (PMA) despite the loss of PMA binding sites by down regulation.

The tumor promoter phorbol 12-myristate 13-acetate (PMA) reversibly inhibits hexamethylene bisacetamide-induced terminal differentiation of Friend erythroleukemia cells (FELC). We were successful in continuously inhibiting FELC differentiation by PMA up to 125 weeks (about 240 serial passages of cells in the presence of PMA). During that period, FELC can be induced to differentiate and enter terminal cell division upon removal of PMA. PMA-mediated suppression of FELC differentiation was associated with only a low level of globin mRNA accumulation. However, a rapid accumulation of globin mRNA in the cytoplasm followed by hemoglobin accumulation occurred upon removal of PMA. A specific, saturable, high-affinity receptor for phorbol esters is present in FELC, as was shown by binding studies with [3H]phorbol 12,13-dibutyrate. A significant (80%) loss in the number of phorbol ester receptors of FELC was observed after a continuous inhibition of differentiation by PMA for as much as 125 weeks. Despite such a down regulation of phorbol ester receptors, these cells respond to PMA with a dose-response similar to that of their parent cells, which have the normal number of phorbol ester receptors. Thus, PMA can suppress reversibly the accumulation of globin-specific mRNA and terminal differentiation of FELC during prolonged periods, despite loss of receptor sites, and our results suggest that only few phorbol ester receptors may be necessary for complete inhibition of FELC differentiation by PMA.