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Stiffened Extracellular Matrix and Signaling from Stromal Fibroblasts via Osteoprotegerin Regulate Tumor Cell Invasion in a 3-D Tumor in Situ Model

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Cancer Microenvironment

Abstract

Several changes have been described in the stroma surrounding a tumor, including changes in cellular composition, altered extracellular matrix composition and organization, and increases in stiffness. Tumor cells are influenced by the composition, organization, and mechanical properties of the microenvironment, and by signals from stromal cells. Here we sought to test whether signaling from stromal fibroblasts and/or the small change in stiffness observed in vivo surrounding epithelial tumors regulates tumor cell invasion from a model of a tumor in situ. We generated a novel tumor in situ model system in which a tumor spheroid is encased within a collagen-IV containing membrane and further encased within a collagen-I matrix of in vivo stiffness with or without fibroblasts. Effects of the matrix, fibroblasts or fibroblast signals were determined by observing the invasion of tumor cells into the matrix. Effects of reciprocal tumor cell signaling upon fibroblasts were determined by observing markers of fibroblast activation. We found that a stiffened matrix led to increased dissemination of MDA-MB-231 cells from tumor spheroids when no fibroblasts were present and that MCF10A cells maintained a more normal organization with a stiffened matrix. The presence of fibroblasts, or fibroblast conditioned media, attenuated the effect upon MDA-MB-231 cells. We also observed an attenuation of fibroblast activation associated gene expression in the presence of MDA-MB-231 cells, with a paradoxical increase in activation associated contractile activity. Furthermore, we identified osteoprotegerin as a soluble factor released by fibroblasts in the stiffened environment that is key to the inhibition of cell invasion.

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Acknowledgments

We would like to thank Mariah Hahn and Ryan Gilbert as well as members of their labs, Dany Munoz, Jon Zuidema and Chris McKay, for their assistance with biomaterials. This work has been supported by the American Cancer Society Research Scholar Grant (RSG-10-245-01-CSM).

Authors’ Contributions

JSM acquired all of the data and completed the data analysis. JSM and LAL jointly conceived and directed the project, interpreted the data, and drafted and revised the manuscript. Both authors read and approved the final manuscript.

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Authors and Affiliations

  1. Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599-5320, USA

    Joshua S. McLane

  2. Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180-3590, USA

    Joshua S. McLane & Lee A. Ligon

  3. Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, NY, 12180-3590, USA

    Joshua S. McLane & Lee A. Ligon

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  1. Joshua S. McLane
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Correspondence to Lee A. Ligon.

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McLane, J.S., Ligon, L.A. Stiffened Extracellular Matrix and Signaling from Stromal Fibroblasts via Osteoprotegerin Regulate Tumor Cell Invasion in a 3-D Tumor in Situ Model. Cancer Microenvironment 9, 127–139 (2016). https://doi.org/10.1007/s12307-016-0188-z

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