Abstract
Cancers have a complex relationship with the surrounding environment that regulates everything from progression to response to treatment. Cell–cell and cell–matrix interactions are heavily influenced by protease biology. Studies on the tumor microenvironment have revealed a new complexity for proteases, describing novel substrates for classic proteases, and protease-independent roles for these enzymes. The rapid expansion of 3D in vitro model systems provides excellent tools to study the intricate influence of proteases on the tumor microenvironment. Here we describe a spheroid invasion assay, providing a platform to interrogate key protease–matrix interactions in the context of early-stage breast cancer. Incorporation of pharmacological inhibition and RNAi techniques enables the elucidation of key protease-dependent pathways and can be complemented with immunofluorescence analysis to visualize matrix cleavage events and visualize cell behavior during collective cell invasion.
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Acknowledgments
This work was supported by Cancer Research UK (CRUK) (C10847/A27781), Breast Cancer Now (2017NovPR988), Barts Charity (MRC0173), Rosetrees Trust (M501-F1), and CRUK Microscopy Core Service Grant (C16420/A18066) and by a CRUK UK Centre Grant to Barts Cancer Institute (C355/A25137). We would like to dedicate this manuscript in memory of Professor Ulrich auf dem Keller, a wonderful collaborator and inspiring scientist.
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Gibson, S.V., Carter, E.P., Grose, R.P. (2024). Interrogating the Impact of Protease Activity on Tumor Progression Using 3D Spheroid Models. In: Santamaria, S. (eds) Proteases and Cancer. Methods in Molecular Biology, vol 2747. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3589-6_15
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DOI: https://doi.org/10.1007/978-1-0716-3589-6_15
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