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Gene expression profiles of subepithelial myofibroblasts from mouse ileum or colon, implanted together with human cancer cells in mouse xenografts

ABSTRACT: We isolated myofibroblasts from C57BL6/J mouse ileum or colon and analyzed gene expression profiles from the cultured cells. We also isolated human colon cancer cells from xenografts interacting with mouse intestinal subepithelial myofibroblasts (ISEMFs) and analyzed gene expression in sorted human cancer cells. We used Affymetrix microarrays to detail the global gene expression prolfiles in mouse fibroblasts and human colon cancer cells and identified distinct biological processes of differentially expressed genes in mouse fibroblasts and interacting human colon cancer cells

ORGANISM(S): Mus musculus Homo sapiens

PROVIDER: GSE27564 | GEO | 2012/02/24

SECONDARY ACCESSION(S): PRJNA138401

REPOSITORIES: GEO

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Project description:Pericryptal myofibroblasts in the colon and rectum play an important role in regulating the normal colorectal stem cell niche and facilitating tumour progression. Myofibroblasts have previously mostly been distinguished from normal fibroblasts only by the expression of Î± smooth muscle actin (Î±SMA). We now identify AOC3, a surface monoamine oxidase, as a new marker of myofibroblasts by showing that it is the target protein of the myofibroblast reacting monoclonal antibody (mAb), PR2D3. The normal and tumour tissue distribution and the cell line reactivity of AOC3 match that expected for myofibroblasts. We have shown that the surface expression of AOC3 is sensitive to digestion by trypsin and collagenase and that anti-AOC3 antibodies can be used for FACS sorting of myofibroblasts obtained by non-enzymatic procedures. Whole genome microarray mRNA expression profiles of myofibroblasts and skin fibroblasts revealed four additional genes that are significantly expressed differentially between these two cell types; NKX2-3 and LRRC17 are expressed in myofibroblasts and SHOX2 and TBX5 in skin fibroblasts. Transforming Growth Factor Î² (TGFÎ²) substantially down-regulated AOC3 expression in myofibroblasts but not in skin fibroblasts, in which it dramatically increased the expression of Î±SMA. A knockdown of NKX2-3 in myofibroblasts caused a decrease of myofibroblast-related gene expression and an increased expression of the fibroblast associated gene, SHOX2, suggesting that NKX2-3 is a key mediator for maintaining myofibroblast characteristics. Our results show that colorectal myofibroblasts, as defined by the expression of AOC3, NKX2-3 and other markers, are a distinctly different cell type from TGFÎ² activated fibroblasts. colorectal myofibroblast specific markers and expression profiles were sought by comparing four primary myofibroblast cultures to a panel of four dermal and foreskin fibroblast cell lines Four primary myofibroblast cultures established from adult human colon compared to four skin fibroblast cell lines to identify intestinal myofibroblast specific markers

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Gene expression profiles of subepithelial myofibroblasts from mouse ileum or colon, implanted together with human cancer cells in mouse xenografts

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