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Sci. Signal., 3 July 2012
Vol. 5, Issue 231, p. rs5
[DOI: 10.1126/scisignal.2002677]


Quantitative Imaging of Epithelial Cell Scattering Identifies Specific Inhibitors of Cell Motility and Cell-Cell Dissociation

Dinah Loerke1*{dagger}, Quint le Duc2*, Iris Blonk2, Andre Kerstens1{ddagger}, Emma Spanjaard2, Matthias Machacek1§, Gaudenz Danuser1||, and Johan de Rooij1,2||

1 Department of Cell Biology, Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
2 Hubrecht Institute for Developmental Biology and Stem Cell Research, Uppsalalaan 8, 3584 CT Utrecht, Netherlands.

* These authors contributed equally to this work.

{dagger} Present address: Department of Physics and Astronomy, University of Denver, 2112 East Wesley Avenue, Denver, CO 80208, USA.

{ddagger} Present address: Penguin Computing Inc., 45800 Northport Loop West, Fremont, CA 94538, USA.

§ Present address: Novartis Pharma AG, Lichtstrasse 35, CH-4056 Basel, Switzerland.

Present address: Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.

Abstract: The scattering of cultured epithelial cells in response to hepatocyte growth factor (HGF) is a model system that recapitulates key features of metastatic cell behavior in vitro, including disruption of cell-cell adhesions and induction of cell migration. We have developed image analysis tools that do not require fluorescence tagging and that automatically track and characterize three aspects of scattering in live cells: increase in cell motility, loss of cell-cell adhesion, and spatial dispersion of cells (the redistribution of cells during scattering). We used these tools to screen a library of drugs, and we identified several efficient inhibitors of scattering, which we classified as selective inhibitors of either motility or loss of cell-cell adhesion, or as nonselective inhibitors. We validated the inhibitors and putative targets from this screen in two unrelated model cell lines. Using pharmacological treatments and RNA interference (RNAi), we found that nonsteroidal anti-inflammatory drugs inhibited cell-cell dissociation, that indirubins inhibited cell motility, and that cyclin-dependent kinase 1 and ribosomal S6 kinase were signaling intermediates in HGF-induced cell scattering. This assay is suitable for larger-scale screenings of chemical compounds or RNAi libraries.

|| To whom correspondence should be addressed. E-mail: j.derooij{at} (J.d.R.); Gaudenz_Danuser{at} (G.D.)

Citation: D. Loerke, Q. le Duc, I. Blonk, A. Kerstens, E. Spanjaard, M. Machacek, G. Danuser, J. de Rooij, Quantitative Imaging of Epithelial Cell Scattering Identifies Specific Inhibitors of Cell Motility and Cell-Cell Dissociation. Sci. Signal. 5, rs5 (2012).

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