title: The F-actin and adherence-dependent mechanical differentiation of normal epithelial cells after TGF-β1-induced EMT (tEMT) using a microplate measurement system.
abstract: The epithelial to mesenchymal transition (EMT) is known to involve several physiological and pathological phenomena. In this study, we utilized a microplate measurement system (MMS) approach based on the deflection of a flexible micro-cantilever to measure cell stiffness (in Pa) and adhesion force (in nN) of a single cell during EMT with nN resolution. Our results demonstrated that after transforming growth factor-β1 (TGF-β1) induced EMT (tEMT), NMuMG cells became stiffer due to thicker and more abundant F-actin and displayed stronger vinculin accumulation after long-term cell-substrate adhesion. The MMS could distinguish differences in compressive stiffness (219 ± 10 and 287 ± 14 Pa), tensile stiffness (114 ± 14 and 132 ± 12 Pa), and adhesion force (150 ± 42 and 192 ± 31 nN) between cells before and after tEMT. However, without proper development of the F-actin structure and adequate adherent time, the mechanical differences were diminished. After tEMT, the cells with increased stiffness and a cell-substrate adhesion force benefited by migrating more rapidly and had more invasiveness. Thus, this technology has the potential to benefit research focused on cancer diagnosis, drug development, and cell-substrate interactions.
authors.affiliation: Department of Biomedical Engineering, National Cheng Kung University, No.1 University Road, Tainan City, 701, Taiwan.
authors.forename: T H
authors.initials: TH
authors.lastname: Wu
authors.forename: Y W
authors.initials: YW
authors.lastname: Chiou
authors.forename: W T
authors.initials: WT
authors.lastname: Chiu
authors.forename: M J
authors.initials: MJ
authors.lastname: Tang
authors.forename: C H
authors.initials: CH
authors.lastname: Chen
authors.forename: Ming-Long
authors.initials: ML
authors.lastname: Yeh
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D001696:Biomechanical Phenomena; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D002465:Cell Movement / Q000187:drug effects; D015638:Cytochalasin D / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D055595:Mechanical Phenomena; D051379:Mice; D055616:Microtechnology / Q000295:instrumentation*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D016596:Vinculin / Q000378:metabolism
chemical_list: D000199:Actins; D015820:Cadherins; D053773:Transforming Growth Factor beta1; D016596:Vinculin; D015638:Cytochalasin D