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Gene expression profiling of Caco-2 BBe cells suggests a role for specific signaling pathways during intestinal differentiation

¹ Interdepartmental Nutrition Program, Purdue University, West Lafayette 47907
² Department of Statistics, Purdue University, West Lafayette 47907
³ Center for Medical Genomics, Indiana University School of Medicine, Indianapolis, Indiana 46202

We examined the pattern of gene expression resulting from spontaneous differentiation of Caco-2 BBe cells to gain insight into the molecular changes necessary for enterocyte differentiation. RNA was prepared from cells harvested at three cell stages: proliferating (50% confluent, 2 days in culture), postproliferative nondifferentiated (8 days), and differentiated (15 days). Gene expression profiles were determined using Affymetrix Human Genome U95A GeneChips. Differentially expressed genes were identified following statistical analysis (i.e., ANOVA, bootstrapping adjustments to P values, false detection rate criterion). We identified 1,150 unique genes as differentially expressed; expression of 48.6% fell and 46% increased from 2 to 15 days, while 5.4% had expression that either peaked or dipped at 8 days. Genes expressed during differentiation included several small-intestine-specific genes involved in nutrient transport/metabolism, e.g., DCT1, hephaestin, folate receptor 1, sucrase-isomaltase, and apolipoproteins CI, CIII, B100, H, and M, indicating that this colonic adenocarcinoma cell line has a hybrid colonocyte/enterocyte phenotype. Patterns of gene expression based upon functional classification suggest a role for cell-cell/cell-matrix interactions, suppression of Wnt signaling, and activation of TGFß and phosphatidylinositol 3-kinase pathways during enterocyte differentiation.

microarray; Wnt; transforming growth factor ß; phosphatidylinositol 3-kinase

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