Development of EMT Model, and Inhibitor Screening
EMT (Epithelial-Mesenchymal Transition) is a procedure where epithelia cell loses its cell polarity or cell-to-cell adhesion with surrounding cells, and becomes mesenchymal like cells by obtaining capabilities for migration, invasion. When human lung cancer cell line A549 is stimulated by TGF-β in the early stage of forming spheroid, it induces EMT. As a result, spheroids gradually collapse. This morphology changes are observed by time-lapse imaging. In EMT inhibitor screening experiment, degree of the spheroid collapsing is evaluated by the signal of Hypoxia Probe LOX-1. The above phenotypic screening permits development of EMT inhibitors which work as therapeutics agents of cancer invasion and/or metastasis.
Spheroid morphology in EMT model
TGF-β untreated well showed intact cell spheroids while cell spheroids collapsed in TGF-β treated well. Positive EMT inhibitor repelled spheroids collapsing. The degree of EMT was evaluated by measuring of signal intensity of Hypoxia Probe.