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In Vitro Methods to Study the Modulation of Migration and Invasion by Sphingosine-1-Phosphate

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Sphingosine-1-Phosphate

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1697))

Abstract

Sphingosine-1-phosphate (S1P) is a bioactive lipid that modulates migratory behavior of cells during embryonic development. In addition, S1P might promote tumor progression by enhancing migratory ability and invasiveness of tumor cells. Migration is a complex process that implies cytoskeletal reorganization and formation of structures that enable cell movement. Besides having similar requirements than migration, invasion also involves proteolytic degradation of extracellular matrix (ECM). Matrix metalloproteases (MMPs) have been identified to break down components of the ECM, allowing cancer cells to spread out of the primary tumor. In this chapter, we will describe different techniques to study migration and invasion induced by S1P. To this end, we include detailed protocols of end-point assays to study migration/invasion, and zymography assay to analyze MMP-2 and MMP-9 activity that were standardized in our laboratory in human melanoma cell lines.

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Acknowledgments

We thank Y. Ohta and T. Stossel (Brigham and Women’s Hospital) for the M2 cells and Pablo L. Bergami for the Lu1205 and SkMel2 cells. This work was supported by grants from ANPCyT-Argentina PICT 1659-2010 and 2378-2014 (S. Alvarez).

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Author notes

  1. Melina G. Castro and Ludmila E. Campos contributed equally to this work.

Authors and Affiliations

  1. Instituto Multidisciplinario de Investigaciones Biológicas San Luis (IMIBIO-SL) CONICET and Universidad Nacional de San Luis, San Luis, Argentina

    Melina G. Castro, Ludmila E. Campos, Yamila I. Rodriguez & Sergio E. Alvarez

Authors
  1. Melina G. Castro
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  2. Ludmila E. Campos
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  3. Yamila I. Rodriguez
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  4. Sergio E. Alvarez
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Corresponding author

Correspondence to Sergio E. Alvarez .

Editor information

Editors and Affiliations

  1. Centre for Eye Research Australia, The University of Melbourne, East Melbourne, Victoria, Australia

    Alice Pébay

  2. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada

    Kursad Turksen

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Castro, M.G., Campos, L.E., Rodriguez, Y.I., Alvarez, S.E. (2017). In Vitro Methods to Study the Modulation of Migration and Invasion by Sphingosine-1-Phosphate. In: Pébay, A., Turksen, K. (eds) Sphingosine-1-Phosphate. Methods in Molecular Biology, vol 1697. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_51

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  • DOI: https://doi.org/10.1007/7651_2017_51

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7412-2

  • Online ISBN: 978-1-4939-7413-9

  • eBook Packages: Springer Protocols

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