Breast cancer is the second leading cause of cancer-related deaths among women in the United States. Approximately 180,000 new cases of breast cancer are diagnosed each year and a quarter of these are fatal. Early detection is a key to survival of these patients. Unfortunately, no definitive markers are available to diagnose breast cancer at early stages. Identification of such early markers, therefore, is an important priority in breast cancer research. In order to identify early markers, we have focussed on understanding the molecular mechanisms that can lead to conversion of the normal mammary epithelial cells into precancerous immortal cells. Over last several years, we have developed in vitro models of human mammary epithelial cell immortalization which have allowed us to invoke the critical roles of the known tumor suppressor pathways in the maintenance of the untransformed state of mammary epithelial cells. These models are now being used to identify novel genes whose expression is important for normal mammary epithelial cell growth and whose altered expression contributes to breast cell transformation. Characterization of the molecular machinery whose alterations result in early preneoplastic transformation should help identify candidate genes for evaluation as potential early diagnostic markers.