title: 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.
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.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Schwartz
authors.affiliation: College of Medicine, Texas A&M Health Science Center, Bryan, Texas, United States of America.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Rogers
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado, United States of America.
authors.forename: Samir P
authors.initials: SP
authors.lastname: Singh
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado, United States of America.
authors.forename: Justin Y
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors.forename: Samuel G
authors.initials: SG
authors.lastname: Loveland
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors.forename: Justin T
authors.initials: JT
authors.lastname: Koepsel
authors.affiliation: Department of Cancer Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, , United States of America.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Witze
authors.affiliation: 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.
authors.forename: Sara I
authors.initials: SI
authors.lastname: Montanez-Sauri
authors.affiliation: 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.
authors.forename: Kyung E
authors.initials: KE
authors.lastname: Sung
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado at Boulder, Boulder, Colorado, United States of America.
authors.forename: Emi Y
authors.initials: EY
authors.lastname: Tokuda
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, Massachusetts, United States of America.
authors.forename: Yasha
authors.initials: Y
authors.lastname: Sharma
authors.affiliation: Department of Chemical and Materials Engineering, University of Dayton, Dayton, Ohio, United States of America.
authors.forename: Lydia M
authors.initials: LM
authors.lastname: Everhart
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors.forename: Eric H
authors.initials: EH
authors.lastname: Nguyen
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, Massachusetts, United States of America.
authors.forename: Muhammad H
authors.initials: MH
authors.lastname: Zaman
authors.affiliation: 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.
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
authors.affiliation: 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.
authors.forename: Natalie G
authors.initials: NG
authors.lastname: Ahn
authors.affiliation: 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.
authors.forename: William L
authors.initials: WL
authors.lastname: Murphy
authors.affiliation: 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.
authors.forename: Kristi S
authors.initials: KS
authors.lastname: Anseth
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D051929:CD146 Antigen / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D002471:Cell Transformation, Neoplastic; D005109:Extracellular Matrix / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D015870:Gene Expression; D006801:Humans; D020100:Hydrogels; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000737:chemistry; D018716:Molecular Mimicry; D010641:Phenotype; D061251:Primary Cell Culture; D016596:Vinculin / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D051929:CD146 Antigen; D020100:Hydrogels; D019012:Integrin beta1; C497902:MCAM protein, human; C488955:VCL protein, human; D016596:Vinculin; D020782:Matrix Metalloproteinases