Netrin-1 regulates invasion and migration of mouse mammary epithelial cells overexpressing Cripto-1 in vitro and in vivo

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.

Topography of Thalamic Projections Requires Attractive and Repulsive Functions of Netrin-1 in the Ventral Telencephalon

“…We used a commercially available polyclonal antibody initially raised against the extracellular domain of Unc5H1 (anti-rat Unc5H1, R&D Systems) and reported to act as a function-blocking reagent against both Unc5A and Unc5C (Unc5H1 and Unc5H3, respectively [36]). We verified the cross-reactivity of this anti-rat Unc5H1 antibody with mouse Unc5A, 5B, and 5C proteins using a biochemical approach (see Figure S12).…”

Mena deficiency delays tumor progression and decreases metastasis in polyoma middle-T transgenic mouse mammary tumors

“…Interestingly, during nervous system development, Ena/VASP proteins are required for normal response to the axon guidance factors SLIT and Netrin [30,59]. Both SLIT and Netrin have been shown to play an important role in mammary gland morphogenesis [32,60,61] and in regulation of cell invasion across basement membranes [62]. Thus, we speculate that Mena's affects on invasion during development and metastasis may involve SLIT and Netrin.…”

Netrin-1 Regulates Th1/Th2/Th17 Cytokine Production and Inflammation through UNC5B Receptor and Protects Kidney against Ischemia–Reperfusion Injury

“…18 h before renal pedicle clamping. The dose of UNC5B Ab and netrin-1 was chosen based on previous studies (20,(23)(24)(25). Plasma and kidneys obtained from mice at different time intervals after reperfusion were used to determine the extent of inflammation and kidney injury for different treatments.…”