THE INDUCTION OF APOPTOSIS IN HUMAN MAMMARY LUMINAL EPITHELIAL-CELLS BY EXPRESSION OF ACTIVATED C-NEU AND ITS ABROGATION BY GLUCOCORTICOIDS

The effects of expressing neu-T, a mutated constitutively activated form of c-neu, have been examined in the non-transformed conditionally immortalised human mammary luminal epithelial cell line, HB4a. A variant cell line, N4.1, which expressed neu-T, showed evidence of transformation, including partial loss of growth factor dependence and acquisition of anchorage-independent growth, but failed to give rise to tumours in nude mice, indicating that expression of neu-T alone was probably insufficient to cause tumorigenic progression to a full malignant phenotype. During characterisation of the N4.1 cell line, it was observed that under conditions of serum deprivation, it underwent apoptotic cell death, as demonstrated by light microscopy, flow cytometry and DNA gel electrophoresis. The induction of apoptotic cell death in the N4.1 cell line by serum deprivation was abrogated specifically by the addition of steroids with glucocorticoid activity but not any peptide growth factors studied. This study shows the induction of apoptosis by serum deprivation, and its abrogation by glucocorticoids occurring in human mammary luminal epithelial cells transformed by expression of neu-T, and implicates the involvement of receptor protein tyrosine kinases in an apoptotic signalling pathway in this cell type.