Abstract
Hypoxia facilitates tumor invasion and metastasis by promoting neovascularization and co-option of tumor cells in the peritumoral vasculature, leading to dissemination of tumor cells into the circulation. However, until recently, animal models and imaging technology did not enable monitoring of the early events of tumor cell invasion and dissemination in living animals. We recently developed a zebrafish metastasis model to dissect the detailed events of hypoxia-induced tumor cell invasion and metastasis in association with angiogenesis at the single-cell level. In this model, fluorescent DiI-labeled human or mouse tumor cells are implanted into the perivitelline cavity of 48-h-old zebrafish embryos, which are subsequently placed in hypoxic water for 3 d. Tumor cell invasion, metastasis and pathological angiogenesis are detected under fluorescent microscopy in the living fish. The average experimental time for this model is 7 d. Our protocol offers a remarkable opportunity to study molecular mechanisms of hypoxia-induced cancer metastasis.
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Acknowledgements
Y.C.'s laboratory was supported by research grants from the Swedish Research Council, the Swedish Cancer Foundation, the Karolinska Institute Foundation, the Karolinska Institute distinguished professor award, the European Union Integrated Project of Metoxia (Project no. 222741) and the European Research Council (ERC) advanced grant ANGIOFAT (Project no 250021). Z.C. is supported by the Swedish Heart and Lung Foundation.
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Y.C. designed the study. L.D.J., P.R. and Z.C. performed the experiments. L.D.J., P.R., Z.C. and Y.C. analyzed the data. K.H., T.L., J.F.S. and E.W. participated in designing and discussing this study. L.D.J., P.R., Z.C. and Y.C. wrote the paper.
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Rouhi, P., Jensen, L., Cao, Z. et al. Hypoxia-induced metastasis model in embryonic zebrafish. Nat Protoc 5, 1911–1918 (2010). https://doi.org/10.1038/nprot.2010.150
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DOI: https://doi.org/10.1038/nprot.2010.150
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