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The Z-cad dual fluorescent sensor detects dynamic changes between the epithelial and mesenchymal cellular states
M J Toneff1, A Sreekumar1, A Tinnirello2
1Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA.
BMC Biology
|June 19, 2016
Summary
A new Z-cad dual sensor tracks epithelial to mesenchymal transition (EMT) in live cancer cells. This tool effectively identifies cancer stem cell (CSC) properties and dynamic cellular changes during EMT and MET.
Area of Science:
- * Cancer Biology
- * Cellular Plasticity
- * Molecular Oncology
Background:
- * Epithelial to mesenchymal transition (EMT) is crucial in carcinoma metastasis, therapy resistance, and cancer stem cell (CSC) properties.
- * Current methods for detecting EMT/CSC phenotypes (cell surface antigens, Cre recombination, immunostaining) are limited as they cannot track dynamic changes in live cells.
- * Live-cell fluorescent sensors tracking EMT regulators are needed to study dynamic cellular plasticity in cancer and development.
Purpose of the Study:
- * To develop a novel fluorescent reporter system for real-time monitoring of EMT and mesenchymal to epithelial transition (MET).
- * To enable the detection and isolation of subpopulations of cells undergoing EMT/MET.
- * To improve the study of cellular plasticity in cancer pathogenesis and normal development.
Main Methods:
- * Development of a lentiviral-based, dual fluorescent reporter system (Z-cad dual sensor).
- * The Z-cad sensor comprises destabilized green fluorescent protein (linked to ZEB1 3' UTR) and red fluorescent protein (driven by E-cadherin promoter).
- * Detection of EMT/MET using flow cytometry and fluorescence microscopy in breast cancer cells.
Main Results:
- * The Z-cad dual sensor successfully detected EMT and MET in breast cancer cells.
- * Dynamic changes in cellular populations during MET were observed.
- * The sensor effectively identified and isolated mesenchymal subpopulations and cells with CSC-like properties, outperforming single-reporter sensors.
Conclusions:
- * The Z-cad dual sensor accurately reports epithelial/mesenchymal state by monitoring key regulators.
- * Live-cell imaging of dynamic EMT/MET fluctuations offers advantages over existing methods for studying cellular plasticity.
- * The Z-cad sensor is versatile and adaptable for in vitro and in vivo studies of EMT/MET in normal and disease states.
