title: Analysis of Epithelial-Mesenchymal Transition Induced by Transforming Growth Factor β.
abstract: In recent years, the importance of the cell biological process of epithelial-mesenchymal transition (EMT) has been established via an exponentially growing number of reports. EMT has been documented during embryonic development, tissue fibrosis, and cancer progression in vitro, in animal models in vivo and in human specimens. EMT relates to many molecular and cellular alterations that occur when epithelial cells undergo a switch in differentiation that generates mesenchymal-like cells with newly acquired migratory and invasive properties. In addition, EMT relates to a nuclear reprogramming similar to the one occurring in the generation of induced pluripotent stem cells. Via such a process, EMT is gradually established to promote the generation and maintenance of adult tissue stem cells which under disease states such as cancer, are known as cancer stem cells. EMT is induced by developmental growth factors, oncogenes, radiation, and hypoxia. A prominent growth factor that causes EMT is transforming growth factor β (TGF-β).A series of molecular and cellular techniques can be applied to define and characterize the state of EMT in diverse biological samples. These methods range from DNA and RNA-based techniques that measure the expression of key EMT regulators and markers of epithelial or mesenchymal differentiation to functional assays of cell mobility, invasiveness and in vitro stemness. This chapter focuses on EMT induced by TGF-β and provides authoritative protocols and relevant reagents and citations of key publications aiming at assisting newcomers that enter this prolific area of biomedical sciences, and offering a useful reference tool to pioneers and aficionados of the field.
authors.affiliation: Inserm U1052, Centre de Recherche en Cancérologie de Lyon, 69000, Lyon, France.|CNRS UMR 5286, Centre de Recherche en Cancérologie de Lyon, 69000, Lyon, France.|Université de Lyon, 69000, Lyon, France.|Université Lyon 1, 69000, Lyon, France.|Centre Léon Bérard, 69000, Lyon, France.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Valcourt
authors.affiliation: Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, Box 595, 751 24, Uppsala, Sweden.
authors.forename: Jonathon
authors.initials: J
authors.lastname: Carthy
authors.affiliation: Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, Box 595, 751 24, Uppsala, Sweden.|Department of Experimental Pathology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Okita
authors.affiliation: Inserm U1052, Centre de Recherche en Cancérologie de Lyon, 69000, Lyon, France.|CNRS UMR 5286, Centre de Recherche en Cancérologie de Lyon, 69000, Lyon, France.|Université de Lyon, 69000, Lyon, France.|Université Lyon 1, 69000, Lyon, France.|Centre Léon Bérard, 69000, Lyon, France.
authors.forename: Lindsay
authors.initials: L
authors.lastname: Alcaraz
authors.affiliation: Department of Experimental Pathology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
authors.forename: Mitsuyasu
authors.initials: M
authors.lastname: Kato
authors.affiliation: INSERM UMR 911 CRO2, Faculty of Pharmacy, Marseille, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Thuault
authors.affiliation: Inserm U1052, Centre de Recherche en Cancérologie de Lyon, 69000, Lyon, France.|CNRS UMR 5286, Centre de Recherche en Cancérologie de Lyon, 69000, Lyon, France.|Université de Lyon, 69000, Lyon, France.|Université Lyon 1, 69000, Lyon, France.|Centre Léon Bérard, 69000, Lyon, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Bartholin
authors.affiliation: Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, Box 595, 751 24, Uppsala, Sweden. aris.moustakas@imbim.uu.se.|Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, Box 582, 751 23, Uppsala, Sweden. aris.moustakas@imbim.uu.se.
authors.forename: Aristidis
authors.initials: A
authors.lastname: Moustakas
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D003094:Collagen; D004338:Drug Combinations; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects* / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006801:Humans; D066298:In Vitro Techniques; D007797:Laminin; D051379:Mice; D011509:Proteoglycans; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014945:Wound Healing
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D016212:Transforming Growth Factor beta; C059485:matrigel; D003094:Collagen
keywords: Cellular plasticity; Epithelial–mesenchymal transition; Invasion; Motility; Signal transduction; Transcription factor; Transforming growth factor β