title: Mapping proteolytic processing in the secretome of gastric cancer-associated myofibroblasts reveals activation of MMP-1, MMP-2, and MMP-3.
abstract: Cancer progression involves changes in extracellular proteolysis, but the contribution of stromal cell secretomes to the cancer degradome remains uncertain. We have now defined the secretome of a specific stromal cell type, the myofibroblast, in gastric cancer and its modification by proteolysis. SILAC labeling and COFRADIC isolation of methionine containing peptides allowed us to quantify differences in gastric cancer-derived myofibroblasts compared with myofibroblasts from adjacent tissue, revealing increased abundance of several proteases in cancer myofibroblasts including matrix metalloproteinases (MMP)-1 and -3. Moreover, N-terminal COFRADIC analysis identified cancer-restricted proteolytic cleavages, including liberation of the active forms of MMP-1, -2, and -3 from their inactive precursors. In vivo imaging confirmed increased MMP activity when gastric cancer cells were xenografted in mice together with gastric cancer myofibroblasts. Western blot and enzyme activity assays confirmed increased MMP-1, -2, and -3 activity in cancer myofibroblasts, and cancer cell migration assays indicated stimulation by MMP-1, -2, and -3 in cancer-associated myofibroblast media. Thus, cancer-derived myofibroblasts differ from their normal counterparts by increased production and activation of MMP-1, -2, and -3, and this may contribute to the remodelling of the cancer cell microenvironment.
authors.affiliation: Institute of Translational Medicine, University of Liverpool, Liverpool, UK.
authors.forename: Christopher
authors.initials: C
authors.lastname: Holmberg
authors.forename: Bart
authors.initials: B
authors.lastname: Ghesquière
authors.forename: Francis
authors.initials: F
authors.lastname: Impens
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
authors.forename: J Dinesh
authors.initials: JD
authors.lastname: Kumar
authors.forename: Nicole
authors.initials: N
authors.lastname: Cash
authors.forename: Sandhir
authors.initials: S
authors.lastname: Kandola
authors.forename: Peter
authors.initials: P
authors.lastname: Hegyi
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Wang
authors.forename: Graham J
authors.initials: GJ
authors.lastname: Dockray
authors.forename: Andrea
authors.initials: A
authors.lastname: Varro
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000378:metabolism*; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis / Q000378:metabolism*; D051379:Mice; D058628:Myofibroblasts / Q000378:metabolism* / Q000473:pathology; D059748:Proteolysis; D013274:Stomach Neoplasms / Q000378:metabolism* / Q000473:pathology; D059016:Tumor Microenvironment / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1