Characterization of the Tumor Secretome from Tumor Interstitial Fluid (TIF).
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.
Danish Cancer Society Research Center, Genome Integrity Unit, Cancer Proteomics Group, Starndboulevarden 49, Copenhagen, 2100, Denmark.
Pavel
P
Gromov
Danish Cancer Society Research Center, Genome Integrity Unit, Cancer Proteomics Group, Starndboulevarden 49, Copenhagen, 2100, Denmark.
Irina
I
Gromova
D014408:Biomarkers, Tumor; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D045604:Extracellular Fluid / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology; D020543:Proteome; D040901:Proteomics / Q000379:methods; D059016:Tumor Microenvironment
D014408:Biomarkers, Tumor; D020543:Proteome
2D-PAGE; Proteomic profiling; Tissue secretome; Tissue section; Tumor interstitial fluid; Tumor microenvironment