Maria Magdalena Koczorowska1, Charlotte Friedemann1, Klaus Geiger2,3, Marie Follo2,3, Martin Lothar Biniossek1, Oliver Schilling4,5,6. 1. Institute of Molecular Medicine and Cell Research, University of Freiburg, Stefan-Meier-Str. 17, D-79104, Freiburg, Germany. 2. Lighthouse Core Facility, Zentrum für Translationale Zellforschung (ZTZ), Medical Center - University of Freiburg, Freiburg, Germany. 3. Department of Medicine I, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. 4. Institute of Molecular Medicine and Cell Research, University of Freiburg, Stefan-Meier-Str. 17, D-79104, Freiburg, Germany. oliver.schilling@mol-med.uni-freiburg.de. 5. BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany. oliver.schilling@mol-med.uni-freiburg.de. 6. German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany. oliver.schilling@mol-med.uni-freiburg.de.
Abstract
BACKGROUND: Solid tumors contain various components that together form the tumor microenvironment. Cancer associated fibroblasts (CAFs) are capable of secreting and responding to signaling molecules and growth factors. Due to their role in tumor development, CAFs are considered as potential therapeutic targets. A prominent tumor-associated signaling molecule is transforming growth factor β (TGFβ), an inducer of epithelial-to-mesenchymal transition (EMT). The differential action of TGFβ on CAFs and ETCs (epithelial tumor cells) has recently gained interest. Here, we aimed to investigate the effects of TGFβ on CAFs and ETCs at the proteomic level. METHODS: We established a 2D co-culture system of differentially fluorescently labeled CAFs and ETCs and stimulated this co-culture system with TGFβ. The respective cell types were separated using FACS and subjected to quantitative analyses of individual proteomes using mass spectrometry. RESULTS: We found that TGFβ treatment had a strong impact on the proteome composition of CAFs, whereas ETCs responded only marginally to TGFβ. Quantitative proteomic analyses of the different cell types revealed up-regulation of extracellular matrix (ECM) proteins in TGFβ treated CAFs. In addition, we found that the TGFβ treated CAFs exhibited increased N-cadherin levels. CONCLUSIONS: From our data we conclude that CAFs respond to TGFβ treatment by changing their proteome composition, while ETCs appear to be rather resilient.
BACKGROUND: Solid tumors contain various components that together form the tumor microenvironment. Cancer associated fibroblasts (CAFs) are capable of secreting and responding to signaling molecules and growth factors. Due to their role in tumor development, CAFs are considered as potential therapeutic targets. A prominent tumor-associated signaling molecule is transforming growth factor β (TGFβ), an inducer of epithelial-to-mesenchymal transition (EMT). The differential action of TGFβ on CAFs and ETCs (epithelial tumor cells) has recently gained interest. Here, we aimed to investigate the effects of TGFβ on CAFs and ETCs at the proteomic level. METHODS: We established a 2D co-culture system of differentially fluorescently labeled CAFs and ETCs and stimulated this co-culture system with TGFβ. The respective cell types were separated using FACS and subjected to quantitative analyses of individual proteomes using mass spectrometry. RESULTS: We found that TGFβ treatment had a strong impact on the proteome composition of CAFs, whereas ETCs responded only marginally to TGFβ. Quantitative proteomic analyses of the different cell types revealed up-regulation of extracellular matrix (ECM) proteins in TGFβ treated CAFs. In addition, we found that the TGFβ treated CAFs exhibited increased N-cadherin levels. CONCLUSIONS: From our data we conclude that CAFs respond to TGFβ treatment by changing their proteome composition, while ETCs appear to be rather resilient.
Authors: Julia D Knopf; Stefan Tholen; Maria M Koczorowska; Olivier De Wever; Martin L Biniossek; Oliver Schilling Journal: Biochim Biophys Acta Date: 2015-07-23
Authors: M M Koczorowska; S Tholen; F Bucher; L Lutz; J N Kizhakkedathu; O De Wever; U F Wellner; M L Biniossek; A Stahl; S Lassmann; O Schilling Journal: Mol Oncol Date: 2015-08-11 Impact factor: 6.603
Authors: M G Bachem; E Schneider; H Gross; H Weidenbach; R M Schmid; A Menke; M Siech; H Beger; A Grünert; G Adler Journal: Gastroenterology Date: 1998-08 Impact factor: 22.682