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Quantitative FRET Imaging to Visualize the Invasiveness of Live Breast Cancer Cells

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Title: Quantitative FRET Imaging to Visualize the Invasiveness of Live Breast Cancer Cells
Author(s): Lu, Shaoying; Wang, Yi; Huang, He; Pan, Yijia; Chaney, Eric J.; Boppart, Stephen A.; Ozer, Howard; Strongin, Alex Y.; Wang, Yingxiao
Subject(s): cancer cell collagen degradation cancer invasion
Abstract: Matrix metalloproteinases (MMPs) remodel tumor microenvironment and promote cancer metastasis. Among the MMP family proteases, the proteolytic activity of the pro-tumorigenic and pro-metastatic membrane-type 1 (MT1)-MMP constitutes a promising and targetable biomarker of aggressive cancer tumors. In this study, we systematically developed and characterized several highly sensitive and specific biosensors based on fluorescence resonant energy transfer (FRET), for visualizing MT1-MMP activity in live cells. The sensitivity of the AHLR-MT1-MMP biosensor was the highest and five times that of a reported version. Hence, the AHLR biosensor was employed to quantitatively profile the MT1-MMP activity in multiple breast cancer cell lines, and to visualize the spatiotemporal MT1-MMP activity simultaneously with the underlying collagen matrix at the single cell level. We detected a significantly higher level of MT1-MMP activity in invasive cancer cells than those in benign or non-invasive cells. Our results further show that the high MT1-MMP activity was stimulated by the adhesion of invasive cancer cells onto the extracellular matrix, which is precisely correlated with the cell’s ability to degrade the collagen matrix. Thus, we systematically optimized a FRET-based biosensor, which provides a powerful tool to detect the pro-invasive MT1-MMP activity at single cell levels. This readout can be applied to profile the invasiveness of single cells from clinical samples, and to serve as an indicator for screening anti-cancer inhibitors.
Issue Date: 2013-03
Publisher: Public Library of Science
Citation Info: Lu, S. Y., Wang, Y., Huang, H., Pan, Y. J., Chaney, E. J., Boppart, S. A., Ozer, H., Strongin, A. Y. and Wang, Y. X. Quantitative FRET Imaging to Visualize the Invasiveness of Live Breast Cancer Cells. Plos One. 2013. 8(3). doi: 10.1371/journal.pone.0058569.
Type: Article
Description: Copyright: 2013 Lu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
URI: http://hdl.handle.net/10027/12829
ISSN: 1932-6203
Sponsor: This work is supported by grants from NIH HL098472, CA139272, NS063405, NSF CBET0846429, CMMI0800870 (Yingxiao W. and S.L.), and the UIC Cancer Center Seed Grant (Yingxiao W. and H. O.).
Date Available in INDIGO: 2014-03-18
 

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