Changes in epidermal growth factor receptor and its messenger ribonucleic acid levels in human placenta and isolated trophoblast cells during pregnancy.

Abstract

We measured the amounts of epidermal growth factor receptor (EGFR) in plasma membrane fractions purified from early, middle, and term placentas and from isolated trophoblast cells, and the amounts of EGFR mRNA were measured in whole placentas. Binding studies were performed using [125I] human EGF as ligand; two classes (high and low) of binding sites were found in placental and trophoblast cell plasma membranes. Although dissociation constants (Kd) were not significantly different in placental plasma membranes from the three stages of pregnancy, the number of binding sites increased significantly during pregnancy. The mean numbers of binding sites were 0.72 +/- 0.18 (+/- SE), 1.02 +/- 0.10, and 1.89 +/- 0.21 pmol/mg protein (high affinity sites) in plasma membrane fractions from early, middle, and term whole placentas, respectively. A similar significant increase was found when the membranes were prepared from isolated trophoblast cells. At the same time, total cellular RNA was isolated, denatured, and blotted onto nitrocellulose membranes. Then, hybridization with 32P-labeled pE 7, a cDNA of EGFR, or 32P-labeled v-erb-B oncogene, autoradiography, and densitometry were performed. The EGFR mRNA increased proportionally to the changes in EGFR during pregnancy. These results indicate that EGF-binding sites and EGFR production increase in human placentas throughout the gestational period.