Summary
We describe an in vivo invasion assay that enables the collection of invasive cells from the primary tumor. In addition to determination of the endogenous, unstimulated invasive properties of cells in vivo, the assay can take advantage of the chemotactic properties of cancer cells. Microneedles are filled with a mixture of extracellular matrix components such as Matrigel with or without a chemoattractant such as EGF, and then introduced into the primary tumor of a rat or mouse that is generated either by orthotopic injection of carcinoma cell lines or by a transgene such as polyoma Middle T. Over the course of 4 h the invasive cell population enters the needles while the animal is kept under anesthesia. At the end of the collection time, the invasive cells are extruded from the microneedles and can be analyzed in terms of the number and type of cells that invade in response to defined stimuli. By including pharmacological inhibitors in the needle, signaling pathways contributing to in vivo invasion can also be identified. This assay leads to a better understanding of the cell types and signaling involved in the tumor microenvironment, and has the potential to be applied to a variety of in vivo models.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wells, A. (2000) Tumor invasion: role of growth factor-induced cell motility. Adv. Cancer Res. 78, 31–101.
Smirnova, T., and Segall, J. E. (2007) Amoeboid chemotaxis: future challenges and opportunities. Cell Adh Migr 1, 165–170.
McSherry, E. A., Donatello, S., Hopkins, A. M., and McDonnell, S. (2007) Molecular basis of invasion in breast cancer. Cell Mol. Life Sci. 64, 3201–3218.
Farrow, B., Albo, D., and Berger, D. H. (2008) The role of the tumor microenvironment in the progression of pancreatic cancer. J. Surg. Res. 149, 319–328.
Muller, A., Homey, B., Soto, H., Ge, N., Catron, D., Buchanan, M. E., et al. (2001) Involvement of chemokine receptors in breast cancer metastasis. Nature 410, 50–6.
Kedrin, D., van Rheenen, J., Hernandez, L., Condeelis, J., and Segall, J. E. (2007) Cell motility and cytoskeletal regulation in invasion and metastasis. J. Mammary Gland Biol. Neoplasia 12, 143–152.
Sidani, M., Wyckoff, J., Xue, C., Segall, J. E., and Condeelis, J. (2006) Probing the microenvironment of mammary tumors using multiphoton microscopy. J. Mammary Gland Biol. Neoplasia 11, 151–163.
Goswami, S., Sahai, E., Wyckoff, J. B., Cammer, M., Cox, D., Pixley, F. J., et al. (2005) Macrophages promote the invasion of breast carcinoma cells via a colony-stimulating factor-1/epidermal growth factor paracrine loop. Cancer Res. 65, 5278–5283.
Gaggioli, C., Hooper, S., Hidalgo-Carcedo, C., Grosse, R., Marshall, J. F., Harrington, K., and Sahai, E. (2007) Fibroblast-led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells. Nat. Cell Biol. 9, 1392–1400.
Wyckoff, J. B., Segall, J. E., and Condeelis, J. S. (2000) The collection of the motile population of cells from a living tumor. Cancer Res. 60, 5401–5404.
Wyckoff, J., Wang, W., Lin, E. Y., Wang, Y., Pixley, F., Stanley, E. R., et al. (2004) A paracrine loop between tumor cells and macrophages is required for tumor cell migration in mammary tumors. Cancer Res. 64, 7022–9.
Wang, W., Goswami, S., Lapidus, K., Wells, A. L., Wyckoff, J. B., Sahai, E., et al. (2004) Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors. Cancer Res. 64, 8585–8594.
Wang, W., Wyckoff, J. B., Wang, Y., Bottinger, E. P., Segall, J. E., and Condeelis, J. S. (2003) Gene expression analysis on small numbers of invasive cells collected by chemotaxis from primary mammary tumors of the mouse. BMC Biotechnol. 3, 13.
Condeelis, J., Singer, R. H., and Segall, J. E. (2005) The great escape: when cancer cells hijack the genes for chemotaxis and motility. Annu. Rev. Cell Dev. Biol. 21, 695–718.
Guy, C. T., Cardiff, R. D., and Muller, W. J. (1992) Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol. Cell. Biol. 12, 954–961.
Austyn, J. M., and Gordon, S. (1981) F4/80, a monoclonal antibody directed specifically against the mouse macrophage, Eur. J. Immunol. 11, 805–815.
Acknowledgments
This work was supported by NIH CA110269 (LH), NIH CA100324 (JC, JES, JW), and NIH CA77522 (TS and JES). J.E.S. is the Betty and Sheldon Feinberg Senior Faculty Scholar in Cancer Research. Authors LH and TS contributed equally.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press
About this protocol
Cite this protocol
Hernandez, L., Smirnova, T., Wyckoff, J., Condeelis, J., Segall, J.E. (2009). In Vivo Assay for Tumor Cell Invasion. In: Jin, T., Hereld, D. (eds) Chemotaxis. Methods in Molecular Biology™, vol 571. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-198-1_15
Download citation
DOI: https://doi.org/10.1007/978-1-60761-198-1_15
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-197-4
Online ISBN: 978-1-60761-198-1
eBook Packages: Springer Protocols