1. Netrin-1 regulates invasion and migration of mouse mammary epithelial cells overexpressing Cripto-1 in vitro and in vivo
- Author
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Robert Callahan, Luigi Strizzi, Caterina Bianco, David S. Salomon, Wissam Abdallah, Lindsay Hinck, Morihisa Hirota, Shin Hamada, Nicola Normanno, Dina Raafat, Youping Sun, Cindy M. Chang, and Ahmed Raafat
- Subjects
Genetically modified mouse ,Mammary gland ,Gene Expression ,Mammary Neoplasms, Animal ,Mice, Transgenic ,Receptors, Cell Surface ,Biology ,Cripto ,Cell Line ,Mesoderm ,Mice ,Mammary Glands, Animal ,Cell Movement ,Netrin ,medicine ,Morphogenesis ,Animals ,Humans ,Nerve Growth Factors ,RNA, Messenger ,RNA, Small Interfering ,Protein kinase B ,Membrane Glycoproteins ,Epidermal Growth Factor ,Tumor Suppressor Proteins ,Epithelial Cells ,Cell Biology ,Netrin-1 ,Epithelium ,Cell biology ,Extracellular Matrix ,Neoplasm Proteins ,medicine.anatomical_structure ,Cell culture ,Immunology ,Female ,Signal transduction ,Netrin Receptors ,hormones, hormone substitutes, and hormone antagonists ,Signal Transduction - Abstract
The neuronal guidance molecule, Netrin-1, has been suggested to play a role in the adhesion and migration of the mammary gland epithelium. Human and mouse Cripto-1 induce proliferation, migration, invasion and colony formation by epithelial cells in 3D matrices. Here we investigate whether Netrin-1 affects these Cripto-1-dependent activities in mouse mammary epithelial cells. Overexpression of Cripto-1 in EpH4 and HC-11 cells (EpH4/Cripto-1 or HC-11/Cripto-1) was associated with low expression of Netrin-1 and increased expression of its receptor Neogenin compared to that of wild-type cells. No change was observed in the expression of the other Netrin-1 receptor, UNC5H1. Treating EpH4/Cripto-1 or HC-11/Cripto-1 mammary cells with exogenous soluble Netrin-1 resulted in increased expression of E-cadherin and UNC5H1, decreased expression of vimentin and decreased activation of Akt as determined by western blotting. Colony formation by Eph4/Cripto-1 cells in 3D gels was significantly reduced in proximity to a Netrin-1 source, and mammary glands of transgenic mice overexpressing human Cripto-1 showed altered ductal growth in proximity to implanted Netrin-1-releasing pellets. Terminal end buds in the treated transgenic mice mammary glands also showed increased expression of E-cadherin and UNC5H1 and decreased expression of active Akt determined by immunohistochemistry. Together, these results suggest that regulation of Netrin-1 expression is important in regulating Cripto-1-dependent invasion and migration of mammary epithelial cells.
- Published
- 2005