1. 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
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Schwartz MP, Rogers RE, Singh SP, Lee JY, Loveland SG, Koepsel JT, Witze ES, Montanez-Sauri SI, Sung KE, Tokuda EY, Sharma Y, Everhart LM, Nguyen EH, Zaman MH, Beebe DJ, Ahn NG, Murphy WL, and Anseth KS
- Subjects
- Actins genetics, Actins metabolism, CD146 Antigen genetics, CD146 Antigen metabolism, Cell Adhesion, Cell Culture Techniques, Cell Line, Tumor, Extracellular Matrix chemistry, Fibroblasts metabolism, Gene Expression, Humans, Hydrogels, Integrin beta1 genetics, Integrin beta1 metabolism, Matrix Metalloproteinases chemistry, Molecular Mimicry, Primary Cell Culture, Vinculin genetics, Vinculin metabolism, Cell Movement genetics, Cell Transformation, Neoplastic, Fibroblasts pathology, 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.
- Published
- 2013
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