Abstract
Circulating tumor cells (CTCs) may represent an easily accessible source of tumor material to assess genetic aberrations such as gene-rearrangements or gene-amplifications and screen cancer patients eligible for targeted therapies. As the number of CTCs is a critical parameter to identify such biomarkers, we developed fluorescent in situ hybridization (FISH) for CTCs enriched on filters (filter-adapted-FISH, FA-FISH). Here, we describe the FA-FISH protocol, the combination of immunofluorescent staining (DAPI/CD45) and FA-FISH techniques, as well as the semi-automated microscopy method that we developed to improve the feasibility and reliability of FISH analyses in filtration-enriched CTC.
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References
Farace F, Massard C, Vimond N, Drusch F, Jacques N, Billiot F et al (2011) A direct comparison of CellSearch and ISET for circulating tumour-cell detection in patients with metastatic carcinomas. Br J Cancer 105(6):847–853
Hofman V, Ilie MI, Long E, Selva E, Bonnetaud C, Molina T et al (2011) Detection of circulating tumor cells as a prognostic factor in patients undergoing radical surgery for non-small-cell lung carcinoma: comparison of the efficacy of the CellSearch Assay and the isolation by size of epithelial tumor cell method. Int J Cancer 129(7):1651–1660
Krebs MG, Hou JM, Sloane R, Lancashire L, Priest L, Nonaka D et al (2012) Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches. J Thorac Oncol 7(2):306–315
Pailler E, Adam J, Barthelemy A, Oulhen M, Auger N, Valent A et al (2013) Detection of circulating tumor cells harboring a unique ALK rearrangement in ALK-positive non-small-cell lung cancer. J Clin Oncol 31(18):2273–2281
Pailler E, Auger N, Lindsay CR, Vielh P, Islas-Morris-Hernandez A, Borget I et al (2015) High level of chromosomal instability in circulating tumor cells of ROS1-rearranged non-small-cell lung cancer. Ann Oncol 26(7):1408–1415
Massard C, Oulhen M, Le Moulec S, Auger N, Foulon S, Abou-Lovergne A et al (2016) Phenotypic and genetic heterogeneity of tumor tissue and circulating tumor cells in patients with metastatic castrationresistant prostate cancer: a report from the PETRUS prospective study. Oncotarget 7(34):55069–55082
Pailler E, Oulhen M, Billiot F, Galland A, Auger N, Faugeroux V et al (2016) Method for semi-automated microscopy of filtration-enriched circulating tumor cells. BMC Cancer 16:477
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Oulhen, M., Pailler, E., Faugeroux, V., Farace, F. (2017). Filter-Adapted Fluorescent In Situ Hybridization (FA-FISH) for Filtration-Enriched Circulating Tumor Cells. In: M. Magbanua, M., W. Park, J. (eds) Circulating Tumor Cells. Methods in Molecular Biology, vol 1634. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7144-2_10
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DOI: https://doi.org/10.1007/978-1-4939-7144-2_10
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7143-5
Online ISBN: 978-1-4939-7144-2
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