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PLoS One. 2013 Dec 3;8(12):e81689. doi: 10.1371/journal.pone.0081689. eCollection 2013.

A quantitative comparison of human HT-1080 fibrosarcoma cells and primary human dermal fibroblasts identifies a 3D migration mechanism with properties unique to the transformed phenotype.

Author information

1
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
2
College of Medicine, Texas A&M Health Science Center, Bryan, Texas, United States of America.
3
Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado, United States of America.
4
Department of Cancer Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, , United States of America.
5
Materials Science Program, University of Wisconsin-Madison, Madison, Wisconsin, United States of America ; Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
6
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America ; Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
7
Department of Biomedical Engineering, Boston University, Boston, Massachusetts, United States of America.
8
Department of Chemical and Materials Engineering, University of Dayton, Dayton, Ohio, United States of America.
9
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America ; Paul P. Carbone Comprehensive Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America ; Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
10
Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, Colorado, United States of America ; Howard Hughes Medical Institute, University of Colorado at Boulder, Boulder, Colorado, United States of America.
11
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America ; Department of Orthopedics and Rehabilitation, University of Wisconsin-Madison, Madison, Wisconsin, United States of America ; Materials Science Program, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
12
Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado, United States of America ; Howard Hughes Medical Institute, University of Colorado at Boulder, Boulder, Colorado, United States of America.

Abstract

Here, we describe an engineering approach to quantitatively compare migration, morphologies, and adhesion for tumorigenic human fibrosarcoma cells (HT-1080s) and primary human dermal fibroblasts (hDFs) with the aim of identifying distinguishing properties of the transformed phenotype. Relative adhesiveness was quantified using self-assembled monolayer (SAM) arrays and proteolytic 3-dimensional (3D) migration was investigated using matrix metalloproteinase (MMP)-degradable poly(ethylene glycol) (PEG) hydrogels ("synthetic extracellular matrix" or "synthetic ECM"). In synthetic ECM, hDFs were characterized by vinculin-containing features on the tips of protrusions, multipolar morphologies, and organized actomyosin filaments. In contrast, HT-1080s were characterized by diffuse vinculin expression, pronounced β1-integrin on the tips of protrusions, a cortically-organized F-actin cytoskeleton, and quantitatively more rounded morphologies, decreased adhesiveness, and increased directional motility compared to hDFs. Further, HT-1080s were characterized by contractility-dependent motility, pronounced blebbing, and cortical contraction waves or constriction rings, while quantified 3D motility was similar in matrices with a wide range of biochemical and biophysical properties (including collagen) despite substantial morphological changes. While HT-1080s were distinct from hDFs for each of the 2D and 3D properties investigated, several features were similar to WM239a melanoma cells, including rounded, proteolytic migration modes, cortical F-actin organization, and prominent uropod-like structures enriched with β1-integrin, F-actin, and melanoma cell adhesion molecule (MCAM/CD146/MUC18). Importantly, many of the features observed for HT-1080s were analogous to cellular changes induced by transformation, including cell rounding, a disorganized F-actin cytoskeleton, altered organization of focal adhesion proteins, and a weakly adherent phenotype. Based on our results, we propose that HT-1080s migrate in synthetic ECM with functional properties that are a direct consequence of their transformed phenotype.

PMID:
24349113
PMCID:
PMC3857815
DOI:
10.1371/journal.pone.0081689
[Indexed for MEDLINE]
Free PMC Article

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