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Identification of proteins released by pancreatic cancer cells by multidimensional protein identification technology: a strategy for identification of novel cancer markers
Author(s) -
Mauri Pierluigi,
Scarpa Aldo,
Nascimbeni Anna Chiara,
Benazzi Louise,
Parmagnani Emanuela,
Mafficini Andrea,
Della Peruta Marco,
Bassi Claudio,
Miyazaki Kaoru,
Sorio Claudio
Publication year - 2005
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.04-3000fje
Subject(s) - versican , pancreatic cancer , cancer cell , perlecan , extracellular matrix , cancer , chemistry , cancer research , stroma , biology , biochemistry , microbiology and biotechnology , proteoglycan , immunohistochemistry , immunology , genetics
The purpose of this study is to identify novel proteins released by cancer cells that are involved in extracellular matrix (ECM) remodeling using small‐volume samples and automated technology. We applied multidimensional protein identification technology (MudPIT), which incorporates two‐dimensional capillary chromatography coupled to tandem mass spectrometry to small quantities of serum‐free supernatants of resting or phorbol ester‐activated Suit‐2 pancreatic cancer cells. Selected markers were validated in additional pancreatic cancer cell lines, primary cancers, and xenografted cancer cells. MudPIT analysis of 10 µl of supernatants identified 46 proteins, 21 of which are classified as secreted, and 10 have never been associated with pancreatic cancer. These include CSPG2/versican, Mac25/angiomodulin, IGFBP‐1, HSPG2/perlecan, syndecan 4, FAM3C, APLP2, cyclophilin B, β2 microglobulin, and ICA69. Evidence that cancer cells release these proteins in vivo was obtained for CSPG2/versican and Mac25/angiomodulin by immunohistochemistry on both primary pancreatic cancers and in a model consisting of Suit‐2 cells embedded in an amorphous matrix and implanted in athymic mice. MudPIT allowed efficient and rapid identification of proteins released by cancer cells, including molecules previously undescribed in the type of cancer analyzed. Our finding that pancreatic cancer cells secrete a series of proteoglycans, including versican, perlecan, syndecan 1 and 4, challenges the common view that fibroblasts of tumor stroma are the sole source of these molecules.