Age‐dependent variations of human and rat colon myofibroblasts in culture: Influence on their functional interactions with colon cancer cells

Abstract

Epithelial-mesenchymal interactions play a pivotal role in colon cancer invasion and metastasis. We aimed at elucidating the impact of long-term cultivation on the phenotypic and functional characteristics of primary fibroblasts and their interaction with the human colon adenocarcinoma cell line LoVoC5. We used fibroblasts from human colon tumor tissue, normal human colon mucosa, rat normal colon and 2 rat colon-derived myofibroblast cell lines, MIC316 and MG. The following parameters were studied: cell shape and size, growth curve, intermediate filament expression and extracellular matrix synthesis. Coculture models with or without cell contacts were used to test the effects on LoVoC5 cell proliferation, spreading and adhesion. Irrespective of their origin, fibroblastic cells in primary cultures presented marked phenotypic and functional changes with time. Before passage 5, they presented as large, slow-growing cells expressing vimentin and ␣-smooth muscle actin; synthesizing laminin-1, fibronectin and collagens I and IV; and inducing LoVoC5 proliferation, spreading and adhesion. After passage 15, they presented as small, fast-growing cells inconstantly expressing ␣-smooth muscle actin and synthesizing mainly type I collagen. In coculture with or without cell contacts, they inhibited LoVoC5 proliferation and allowed only limited cell spreading and adhesion. Myofibroblastic cell lines presented as large, fast-growing cells expressing vimentin and ␣-smooth muscle actin and synthesizing mainly type I collagen. They had no significant effects on LoVoC5 proliferation, spreading and adhesion. Our results underline the importance of age-dependent variations in colon mesenchymal cells in culture and for the in vitro study of epithelial-mesenchymal interactions in colon cancer.

Acidic fibroblast growth factor decreases α-smooth muscle actin expression and induces apoptosis in human normal lung fibroblasts

“…Likewise, FGF-1ϩH strongly inhibited (when used simultaneously) and reverted (when used subsequently) the TGF-␤1-induced switching of the human lung fibroblasts to myofibroblasts that was accompanied by a change in cell morphology from a typical flattened, irregular shape characteristic of a myofibroblast-like phenotype to an elongated, spindle-shaped appearance (17,26). The effect of FGF-1ϩH on myofibroblast phenotype was demonstrated by several complementary techniques, including flow cytometry, immunocytochemistry, and quantification of ␣-SMA by real-time RT-PCR and Western blot.…”

“…The effects of FGFs are mediated by binding to the FGF receptor (FGFR) family of receptor tyrosine kinases (19,30,44). Heparin and heparan sulfate glycosaminoglycans act as low-affinity receptors and are part of a dual-receptor system, which, along with a cell surface tyrosine kinase receptor, transduces ligand activation along appropriate cytosolic and nuclear pathways (20,21,26). FGF-1, the first described of this family, regulates cell growth and differentiation and participates in several physiological processes, including angiogenesis and tissue repair (19).…”

Breast fibroblasts modulate epithelial cell proliferation in three-dimensional in vitro co-culture

“…Only early-passage fibroblast cultures are used in experiments to more closely simulate their in vivo phenotype. It has been shown that early-passage (below passage 9) colonic primary fibroblasts maintain many of their in vivo characteristics, including expression of alpha-smooth muscle actin, collagen IV and laminin 1 [28]. Multiple NAF and CAF cultures were utilized because of potential variation among fibroblasts from different individuals and different breast cancers.…”

Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors

“…Cluster II includes many genes that encode secretory proteins and genes involved in cell matrix or matrix modeling (e.g., Fibronectin, TIMP3, ADAMTS1, and TAGLIN). Some of these genes (including Fibronectin and TAGLIN) have been found to be expressed by myofibroblasts as well as smooth muscle cells (11,12). Therefore, we suspect that genes in this cluster most likely represent genes that are expressed by cryptic stromal cells.…”