title: Modeling colon adenocarcinomas in vitro a 3D co-culture system induces cancer-relevant pathways upon tumor cell and stromal fibroblast interaction.
abstract: Activated tumor stroma participates in tumor cell growth, invasion, and metastasis. Normal fibroblasts and cancer-associated fibroblasts (CAFs) have been shown to display distinct gene expression signatures. This molecular heterogeneity may influence the way tumor cells migrate, proliferate, and survive during tumor progression. To test this hypothesis and to better understand the molecular mechanisms that control these interactions, we established a three-dimensional (3D) human cell culture system that recapitulates the tumor heterogeneity observed in vivo. Human colon tumor cells were grown as multicellular spheroids and subsequently co-cultured with normal fibroblasts or CAFs in collagen I gels. This in vitro model system closely mirrors the architecture of human epithelial cancers and allows the characterization of the tumor cell-stroma interactions phenotypically and at the molecular level. Using GeneChip analysis, antibody arrays, and enzyme-linked immunosorbent assays, we demonstrate that the interaction of colon cancer cells with stromal fibroblasts induced different highly relevant cancer expression profiles. Genes involved in invasion, extracellular matrix remodeling, inflammation, and angiogenesis were differentially regulated in our 3D carcinoma model. The modular setup, reproducibility, and robustness of the model make it a powerful tool to identify target molecules involved in signaling pathways that mediate paracrine interactions in the tumor microenvironment and to validate the influence of these molecular targets during tumor growth and invasion in the supporting stroma.
authors.affiliation: Institute of Pathology, Medical University of Vienna, Vienna, Austria. helmut.dolznig@meduniwien.ac.at
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.forename: Christian
authors.initials: C
authors.lastname: Rupp
authors.forename: Christina
authors.initials: C
authors.lastname: Puri
authors.forename: Christian
authors.initials: C
authors.lastname: Haslinger
authors.forename: Norbert
authors.initials: N
authors.lastname: Schweifer
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Wieser
authors.forename: Dontscho
authors.initials: D
authors.lastname: Kerjaschki
authors.forename: Pilar
authors.initials: P
authors.lastname: Garin-Chesa
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002471:Cell Transformation, Neoplastic; D018920:Coculture Techniques; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix; D005347:Fibroblasts / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006801:Humans; D007124:Immunoenzyme Techniques; D008958:Models, Molecular; D020411:Oligonucleotide Array Sequence Analysis; D040081:Protein Array Analysis; D015398:Signal Transduction; D018874:Spheroids, Cellular; D017154:Stromal Cells / Q000473:pathology*; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment
chemical_list: D014408:Biomarkers, Tumor; D016207:Cytokines