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1Departments of Medicine and Cell Biology, University of Alabama at Birmingham, and Veterans Administration Medical Center, and 2Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama 35294; 3Department of Medicine, 4Genomics Institute, Bioinformatics Core, and 5Department of Medicine and Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6144
Submitted 30 July 2003 ; accepted in final form 8 December 2003
Hepatocyte growth factor (HGF) elicits a broad spectrum of biological activities, including epithelial cell dedifferentiation. One of the most widely used and best-studied polarized epithelial cell lines is the Madin-Darby canine kidney (MDCK) cell line. Here, we describe and validate the early response of polarized monolayers of MDCK cells stimulated with recombinant HGF using a novel canine DNA microarray designed to query 12,473 gene sequences. In our survey, eight genes previously implicated in the HGF signaling pathway were differentially regulated, demonstrating that the system was responsive to HGF. Also identified were 117 genes not previously known to be involved in the HGF pathway. The results were confirmed by real-time PCR or Western blot analysis for 38 genes. Of particular interest were the large number of differentially regulated genes encoding small GTPases, proteins involved in endoplasmic reticulum translation, proteins involved in the cytoskeleton, the extracellular matrix, and the hematopoietic and prostaglandin systems.
hepatocyte growth factor; Madin-Darby canine kidney cells
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