Abstract
Circulating tumor cells (CTCs) hold promise as biomarkers to aid in patient treatment stratification and disease monitoring. Because the number of cells is a critical parameter for exploiting CTCs for predictive biomarker's detection, we developed a FISH (fluorescent in situ hybridization) method for CTCs enriched on filters (filter-adapted FISH [FA-FISH]) that was optimized for high cell recovery. To increase the feasibility and reliability of the analyses, we combined fluorescent staining and FA-FISH and developed a semi-automated microscopy method for optimal FISH signal identification in filtration-enriched CTCs . Here we present these methods and their use for the detection and characterization of ALK-, ROS1-, RET-rearrangement in CTCs from non-small-cell lung cancer and ERG-rearrangements in CTCs from prostate cancer patients.
Keywords:
Circulating tumor cells; FA-FISH; Filtration-enrichment; Fluorescent staining; Predictive biomarkers.
Publication types
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Research Support, Non-U.S. Gov't
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Review
MeSH terms
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Anaplastic Lymphoma Kinase
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Carcinoma, Non-Small-Cell Lung* / blood
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Carcinoma, Non-Small-Cell Lung* / genetics
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Female
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Gene Rearrangement*
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Humans
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In Situ Hybridization, Fluorescence / instrumentation
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In Situ Hybridization, Fluorescence / methods
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Lung Neoplasms* / blood
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Lung Neoplasms* / genetics
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Male
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Neoplastic Cells, Circulating / metabolism*
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Prostatic Neoplasms* / blood
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Prostatic Neoplasms* / genetics
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Protein-Tyrosine Kinases* / genetics
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Protein-Tyrosine Kinases* / metabolism
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Proto-Oncogene Proteins c-ret* / genetics
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Proto-Oncogene Proteins c-ret* / metabolism
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Proto-Oncogene Proteins* / genetics
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Proto-Oncogene Proteins* / metabolism
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Receptor Protein-Tyrosine Kinases* / genetics
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Receptor Protein-Tyrosine Kinases* / metabolism
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Transcriptional Regulator ERG / genetics
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Transcriptional Regulator ERG / metabolism
Substances
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ERG protein, human
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Proto-Oncogene Proteins
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Transcriptional Regulator ERG
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ALK protein, human
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Anaplastic Lymphoma Kinase
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Protein-Tyrosine Kinases
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Proto-Oncogene Proteins c-ret
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RET protein, human
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ROS1 protein, human
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Receptor Protein-Tyrosine Kinases