Global expression profiling reveals regulation of CTGF/CCN2 during lactogenic differentiation.

Mammary epithelial cells go through a series of developmental changes during pregnancy and lactation including proliferation, differentiation, secretion and apoptosis. HC11 mouse mammary epithelial cells, which undergo lactogen-induced differentiation in cell culture, were used to follow the changes in gene expression during this process. The expression profiles of over 20,000 genes were compared in HC11 cells undergoing lactogenic differentiation to non-differentiated cells using DNA microarray analysis. Greater than two fold changes were detected in 998 genes in the differentiated cells versus growth controls. Several genes including CTGF/CCN2 exhibited greater than five-fold increase. Validation of the gene expression pattern for more than twenty genes was performed. The results indicate the involvement of numerous genes and pathways in the differentiation of mouse mammary epithelial cells in culture and they identify genetic pathways associated with specific transcriptional regulation. In addition, the expression of a subset of genes regulated by lactogenic differentiation in HC11 cells, including CTGF/CCN2 and osteopontin, was examined in mouse mammary glands revealing expression during pregnancy and lactation that declined during involution of the glands. To probe the mechanism by which epidermal growth factor (EGF), a known inhibitor of lactogenic differentiation in HC11 cells, blocks lactogenesis, the HC11 cells stimulated with lactogenic hormone in the presence of EGF were profiled. This data revealed EGF regulation of a specific subset of genes including important cell cycle regulators. The studies confirm the value of expression profiling in defining gene transcription associated with differentiation of mammary epithelial cells.