Proliferation is required for induction of terminal differentiation of Friend erythroleukemia cells

The relationship between cell proliferation and differentiation has long been a source of controversy. Stimulation of normal erythroid maturation results in a finite number of cell divisions accompanied by a concomitant accumulation of hemoglobin. Friend erythroleukemia cells treated with hexamethylene bisacetamide differentiate in a similar manner, while agents such as hemin apparently induce differentiation without limiting cell proliferation. Aphidicolin, an inhibitor of DNA synthesis, has been reported to induce differentiation in the absence of cell proliferation. Using these three chemicals we have investigated the relationship between cell proliferation and erythrocytic maturation by exposing Friend erythroleukemia cells to either hexamethylene bisacetamide (5 mM), hemin (100 microM), or aphidicolin 1.2 microM) and examining the effects on cell growth, morphology, and hemoglobin production. Proliferation in the presence of hexamethylene bisacetamide is limited to four to five rounds of cell division, while hemin has no inhibitory effect. Hexamethylene bisacetamide initiates the complete erythrocytic maturation program, including cellular structural changes and hemoglobin synthesis. Hemin stimulates only globin gene transcription, not differentiation. Aphidicolin inhibits cell growth within 24 h, but does not induce differentiation. Furthermore, inhibition of proliferation by aphidicolin prevents subsequent hexamethylene bisacetamide induced differentiation. These results indicate that at least one round of cell division is required for initiation of erythrocytic differentiation.