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Epidermal growth factor (EGF) as well as prostaglandin F2 alpha (PGF2 alpha), when added to quiescent, confluent Swiss 3T3 cells, stimulate the initiation of DNA synthesis, which occurs with apparent first-order kinetics after a lag phase of 14-15 hr. These two growth factors appear to stimulate similar events; insulin enhances and hydrocortisone can inhibit the stimulatory effect of either. Here we show that the addition of EGF and PGF2 alpha together, however, results in a synergistic effect seen at the end of the lag phase, but only when EGF and PGF2 alpha are added within 6 hr of each other. Addition of one growth factor 10 or 15 hr after the other delayed the synergy for 15 hr after the addition of the second growth factor. Insulin further increased the rate of entry into the s phage stimulated by EGF and PGF2 alpha together, whereas hydrocortisone inhibited the stimulatory effect observed with either EGF or PGF2 alpha alone. These results suggest that, in spite of the common events responsible for the interactions with the two hormones, EGF and PGF2 alpha must have differences in their sequences of events that initiate DNA synthesis.

Two growth factors and two hormones regulate initiation of DNA synthesis in cultured mouse cells through different pathways of events.