The existence of different classes of EGF receptors in human pancreatic cancer cells has yet not been determined. EGF binding to two cancer cell lines (CAPAN-1 and MIA PaCa-2) was studied. Two classes of EGF binding sites were characterized. The first class of EGF binding sites demonstrated a high affinity and low capacity for EGF, with a Kd of 0.25 +/- 0.11 nM, close to the concentration of EGF suggested to be present in human pancreatic juice. The second class of EGF binding sites had a lower affinity and a higher capacity for EGF, with Kd of 1.78 +/- 0.61 nM. The total number of EGF binding sites was about 40,000/cell. Treatment of the cells with a phorbol ester, TPA, caused a complete loss of the high affinity binding sites and also caused a decrease in the concentration of the lower affinity binding sites present on the cells. Interestingly, with the increasing age of the cells, the concentration of both the high and low affinity EGF binding sites was significantly decreased. In the presence or absence of fetal calf serum, EGF, at concentrations higher than 1.10(-10)M, exerted a dose-dependent mitogenic effect on the growth of the pancreatic cancer cells in culture. These data demonstrate the existence of two classes of binding sites for EGF on some human pancreatic cancer cells and a possible role of EGF in the growth of pancreatic tumors.