Members of the HMG-I/Y family of high-mobility-group chromatin proteins have been demonstrated to regulate gene expression in human cells in vivo. They are thought to function as gene regulatory molecules by acting as architectural transcription factors that modulate DNA and/or chromatin structure. Numerous studies have indicated that elevated HMG-I/Y gene expression is directly correlated with more advanced cancers and with increased metastatic potential. The inducible expression of the HMG-I/Y gene was studied in two human mammary epithelial cell lines, MCF7 and Hs578T, in the presence, or absence, of either 17βdiol or epidermal growth factor (EGF). Northern blot analysis indicated that there was no increase in HMG-I/Y mRNA in the nonmetastatic MCF7 cells when they were treated with either 17βestradiol or EGF. In contrast, in the highly metastatic Hs578T cell line, there is a dramatic induction of HMG-I/Y mRNA expression of up to 23-fold when the cells are treated with EGF. mRNA primer extension analysis indicated that only two (of the possible four different) transcription initiation start sites in the HMG-I/Y gene are induced by EGF treatment of the Hs578T cells. Additional experiments demonstrated that in both epithelial cell types HMG-I/Y mRNAs are very stable (tl/2 of ~30 hr) and that in the Hs578T cells treated with EGF the cellular concentrations of the HMG-I/Y proteins increase concurrently with the induced mRNA levels. Given that HMG-I/Y proteins are regulators of gene activity whose elevated in vivo concentrations are known to be correlated with increased metastatic potential, these data demonstrating an EGF-induced over-expression of HMG-I/Y in the highly metastatic Hs578T, but not in the nonmetastatic MCF7cells, may have important implications concerning the cellular mechanisms involved in the progression of mammary epithelial tumors.