Epidermal growth factor receptor expression on human breast luminal and basal cells in vitro.
The expression of EGF receptors has been studied on luminal and basal cells of human breast in vitro. Primary cultures of normal adult human breast epithelium were prepared as single cell suspensions containing a mixture of luminal and basal cells. The cells were simultaneously immunolabelled with antibodies recognising EMA (luminal epithelial cells), CALLA/CD10 (basal cells) and the epidermal growth factor receptor (EGFR). Flow cytometric analysis of these triple labelled cells detected low levels of EGFR on both cell types, with proportionally more EGFR on basal cells compared with luminal cells. Separated populations of basal and luminal cells were prepared from single cell suspensions by flow sorting or by immunomagnetic methods and cultured with and without EGF. Increased proliferation was detected in both cell types in the presence of EGF. To determine the localisation of the EGF receptor, purified cell populations were immunolabelled with anti-EGFR antibody and an FITC-labelled second antibody for fluorescence light microscopy and colloidal gold-labelled antibody for scanning electron microscopy (SEM). Low levels of EGFR were detected by indirect immunofluorescence on both cell types with higher levels on basal cells compared with luminal cells. The detailed subcellular distribution of the receptor was examined by SEM, with gold-labelling of EGFR detected using a field emission scanning electron microscope with a YAG crystal backscattered electron detector. Both luminal and basal cells expressed EGFR over the upper surface of individual cells when these were growing in isolation, but when cells formed part of a confluent island, levels of EGFR on the upper surface of cells were obviously reduced. Observations made by SEM on cells at the edges of such confluent islands showed that cultured basal cells expressed much higher levels of EGFR on their basal, as compared with their upper surfaces.
Monaghan
P
P
Institute of Cancer Research, Haddow Laboratories, Sutton, Surrey, UK. paulm@icr.ac.uk
Clarke
C L
CL
Perusinghe
N P
NP
Ormerod
M G
MG
O'Hare
M J
MJ
D000293:Adolescent; D000328:Adult; D001940:Breast / Q000737:chemistry*; D002469:Cell Separation; D002478:Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000032:analysis* / Q000096:biosynthesis; D005260:Female; D005434:Flow Cytometry; D017979:Gold Colloid; D006801:Humans; D007150:Immunohistochemistry; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence
D017979:Gold Colloid; D004815:Epidermal Growth Factor; D066246:ErbB Receptors