Epidermal growth factor receptors and EGF-responsiveness of the human breast-carcinoma cell line PMC42

Abstract

PMC42 is an early-passage, well-differentiated, pleiomorphic human breast-cancer cell line. It expresses an average of 2.4 x 10(5) EGF receptor (EGFR) sites per cell when cultures are assayed using a radioreceptor assay. This is a relatively high figure for breast-carcinoma cell lines. Its responses to epidermal growth factor (EGF) were analyzed morphologically and by flow cytometry and immunocytochemistry. PMC42 responded in vitro to EGF with morphological changes, inhibition of "doming" and an increase in DNA synthesis in serum-containing medium in confluent or near-confluent cultures. When EGF receptor sites were localized immunocytochemically in PMC42, they were found to be variably expressed on the membrane of cells grown in the absence of EGF, with discrete areas of strong and weak cells of different morphological appearances; flow cytometry indicated a 10-fold range in EGFR levels between individual cells. Cultures maintained in the presence of EGF were less heterogeneous in their morphology and had a predominantly cytoplasmic, relatively uniform localization of EGFR with down-regulation of membrane EGFR levels. EGF-stimulated cultures were stained simultaneously for the presence of EGFR and the proliferation-related antigen Ki-67 and analyzed by both flow cytometry and immunocytochemistry. No correlation was observed between Ki-67 positivity and level of EGFR expression by individual cells. Electron microscopic localization of receptor sites demonstrated that the receptors were concentrated predominantly on the lateral and basal membranes. These results show that the PMC42 line reflects a number of the properties of heterogeneous breast tumours in situ. It may also continue to express growth-factor receptors in a manner that reflects their topographical distribution on normal luminal breast epithelial cells.