Visualizing Dynamic Changes During TGF-β-Induced Epithelial to Mesenchymal Transition

Abhishek Sinha, Pranav Mehta, Chuannan Fan, Jing Zhang, Dieuwke L. Marvin, Maarten van Dinther, Laila Ritsma, Pouyan E. Boukany, Peter ten Dijke*

*Corresponding author for this work

Research output: Chapter in Book/Conference proceedings/Edited volumeChapterScientificpeer-review

2 Citations (Scopus)
26 Downloads (Pure)

Abstract

Epithelial to mesenchymal transition (EMT) is crucial during embryonic development, tissue fibrosis, and cancer progression. Epithelial cells that display a cobblestone-like morphology can undergo a switch to mesenchymal-like phenotype, displaying an elongated spindle shape or a fibroblast-like morphology. EMT is characterized by timely and reversible alterations of molecular and cellular processes. The changes include loss of epithelial and gain of mesenchymal marker expression, loss of polarity, increased cell migratory and invasive properties. Epithelial cells can progress unevenly during this transition and attain hybrid E/M states or metastable EMT states, referred to as epithelial cell plasticity. To gain a deeper insight into the mechanism of EMT, understanding the dynamic aspects of this process is essential. One of the most prominent factors to induce EMT is the cytokine transforming growth factor-β (TGF-β). This chapter discusses molecular and cellular techniques to monitor TGF-β-induced signaling and EMT changes in normal and cancer cell lines. These methods include measuring the TGF-β-induced activation of its intracellular SMAD effectors proteins and changes in epithelial/mesenchymal marker expression and localization. Moreover, we describe assays of cell migration and dynamic reorganization of the actin cytoskeleton and stress filaments that are frequently part of the TGF-β-induced EMT cellular response.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
EditorsZhike Zi, Xuedong Liu
PublisherHumana Press Inc.
Chapter5
Pages47-65
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2488
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project https://www.openaccess.nl/en/you-share-we-take-care
Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public.

Keywords

  • Cell migration
  • EMP
  • EMT
  • Mesenchymal
  • Metastasis
  • TGF-β

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