We recently reported that the growth of normal rat pleural mesothelial cells (RPMCs) is inhibited by conditioned media from either in vivo or in vitro transformed RPMCs. In this study we report that the growth of normal RPMCs is inhibited by epidermal growth factor (EGF). This was demonstrated by using three methods of investigation. Two types of studies were carried out with growing cells. First, cell counts indicated that the number of cells was reduced in EGF-treated cultures when compared with untreated cultures. Second, the percentage of S cells detected by flow cytometry following treatment with EGF was lower than without EGF. In other experiments, incorporation of tritiated thymidine in confluent cells was decreased by EGF treatment, either in the presence or absence of fetal calf serum; these effects were dose dependent and were observed from 2 ng/ml EGF. Lower EGF concentrations did not significantly modify thymidine incorporation when compared with untreated cells. Analysis of 125I EGF binding experiments by the Scatchard method indicated that RPMCs posses EGF receptors (about 10(5) per cell) with low ligand binding affinity (Kd = 1.7 +/- 0.4 nM). These results indicate that EGF might modulate the growth of RPMCs.