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Characterization of the Tumor Secretome from Tumor Interstitial Fluid (TIF)

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Unconventional Protein Secretion

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1459))

Abstract

Tumor interstitial fluid (TIF) surrounds and perfuses bodily tumorigenic tissues and cells, and can accumulate by-products of tumors and stromal cells in a relatively local space. Interstitial fluid offers several important advantages for biomarker and therapeutic target discovery, especially for cancer. Here, we describe the most currently accepted method for recovering TIF from tumor and nonmalignant tissues that was initially performed using breast cancer tissue. TIF recovery is achieved by passive extraction of fluid from small, surgically dissected tissue specimens in phosphate-buffered saline. We also present protocols for hematoxylin and eosin (H&E) staining of snap-frozen and formalin-fixed, paraffin-embedded (FFPE) tumor sections and for proteomic profiling of TIF and matched tumor samples by high-resolution two-dimensional gel electrophoresis (2D-PAGE) to enable comparative analysis of tumor secretome and paired tumor tissue.

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Acknowledgements

Preparation of this manuscript was supported by the Danish Cancer Society through the Danish Cancer Society Research Center budget and by grants from the John and Birthe Meyer Foundation and EurocanPlatform.

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Authors and Affiliations

  1. Danish Cancer Society Research Center, Genome Integrity Unit, Cancer Proteomics Group, Starndboulevarden 49, Copenhagen, 2100, Denmark

    Pavel Gromov & Irina Gromova

Authors
  1. Pavel Gromov
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  2. Irina Gromova
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Editor information

Editors and Affiliations

  1. Instituto di Bioscienze e Biorisorse, Perugia, Italy

    Andrea Pompa

  2. Instituto di Bioscienze e Biorisorse, Perugia, Italy

    Francesca De Marchis

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Gromov, P., Gromova, I. (2016). Characterization of the Tumor Secretome from Tumor Interstitial Fluid (TIF). In: Pompa, A., De Marchis, F. (eds) Unconventional Protein Secretion. Methods in Molecular Biology, vol 1459. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3804-9_16

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  • DOI: https://doi.org/10.1007/978-1-4939-3804-9_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3802-5

  • Online ISBN: 978-1-4939-3804-9

  • eBook Packages: Springer Protocols

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