Summary
The epithelial to mesenchymal transition (EMT) is the breakdown of epithelial cell morphology that gives way to a more mobile, mesenchymal phenotype. Although this process is fundamental to the development of multicellular organisms, it is also a key occurrence in many diseases, including cancers of epithelial origin E-cadherin is a central component of adherens junctions (AJs), which act as structural and signaling hubs in epithelial cells that oppose EMT. The loss of E-cadherin from the plasma membrane is an early indication of EMT and a marker of poor prognosis in many cancers making the trafficking of E-cadherin an area of great interest. Recent work from the authors’ laboratory has established the role of type Iγ phosphatidylinositol 4-phosphate 5-kinase (PIPKIγ) in the trafficking of E-cadherin by studying the surface accessibility of E-cadherin in endocytosis and recycling assays. Additionally, immunofluorescence data demonstrated that cells lacking PIPKIγ lost E-cadherin at the plasma membrane. The biochemical and microscopic techniques used to investigate the trafficking of E-cadherin are presented herein.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Perez-Moreno, M. and Fuchs, E. (2006) Catenins: keeping cells from getting their signals crossed. Dev. Cell. 11, 601–612.
Weis, W. I. and Nelson, W. J. (2006) Re-solving the cadherin-catenin-actin conundrum. J. Biol. Chem. 281, 35593–35597.
Thiery, J. P. (2003) Epithelial-mesenchymal transitions in development and pathologies. Curr. Opin. Cell Biol. 15, 740–6.
Huber, M. A., Kraut, N., and Beug, H. (2005) Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr. Opin. Cell. Biol. 17, 548–558.
Bremnes, R. M., Veve, R., Hirsch, F. R., and Franklin, W. A. (2002) The E-cadherin cell-cell adhesion complex and lung cancer invasion, metastasis, and prognosis. Lung Cancer. 36, 115–124.
Chen, H. C., Chu, R. Y., Hsu, P. N., et al. (2003) Loss of E-cadherin expression correlates with poor differentiation and invasion into adjacent organs in gastric adenocarcinomas. Cancer Lett. 201, 97–106.
Liu, M., Lawson, G., Delos, M., et al. (2003) Prognostic value of cell proliferation markers, tumour suppressor proteins and cell adhesion molecules in primary squamous cell carcinoma of the larynx and hypopharynx. Eur. Arch. Otorhinolaryngol. 260, 28–34.
D' Souza-Schorey, C. (2005) Disassembling adherens junctions: breaking up is hard to do. Trends Cell Biol. 15, 19–26.
Vasioukhin, V., Bauer, C., Degenstein, L., Wise, B. and Fuchs, E. (2001) Hyperproliferation and defects in epithelial polarity upon conditional ablation of alpha-catenin in skin. Cell 104, 605–617.
Perez-Moreno, M., Davis, M. A., Wong, E., Pasolli, H. A., Reynolds, A. B., and Fuchs, E. (2006) p120-catenin mediates inflammatory responses in the skin. Cell 124, 631–644.
Ling, K., Bairstow, S., Carbonara, C., Turbin, D., Huntsman, D., and Anderson, R. (2007) Type Iγ phosphatidylinositol phosphate kinase modulates adherens junction and E-cadherin trafficking via a direct interaction with μ1B adaptin. J. Cell. Biol. 176, 343–353.
Le, T. L., Yap, A. S., and Stow, J. L. (1999) Recycling of E-cadherin: a potential mechanism for regulating cadherin dynamics. J. Cell. Biol. 146, 219–232.
Sambrook, J. and Russell, D., (2001) Molecular Cloning: A Laboratory Manual. 3 ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Panchuk-Voloshina, N., Haugland, R. P., Bishop-Stewart, J., et al. (1999) Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates. J. Histochem. Cytochem. 47, 1179–1188.
Acknowledgments
The authors would like to thank Nick Schill and Professor Richard Anderson for their editing, advice, and encouragement. This work was supported by R01 GM057549-08 and R01 CA104708-03.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Wagoner, M.P., Ling, K., Anderson, R.A. (2008). Tracking the Transport of E-Cadherin to and From the Plasma Membrane. In: Vancura, A. (eds) Membrane Trafficking. Methods in Molecular Biology, vol 457. Humana Press. https://doi.org/10.1007/978-1-59745-261-8_20
Download citation
DOI: https://doi.org/10.1007/978-1-59745-261-8_20
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-925-3
Online ISBN: 978-1-59745-261-8
eBook Packages: Springer Protocols