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Am J Physiol Renal Physiol (December 9, 2003). doi:10.1152/ajprenal.00270.2003
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Submitted on July 30, 2003
Accepted on December 8, 2003

Gene Expression Alterations During HGF-Induced Dedifferentiation of a Renal Tubular Epithelial Cell Line (MDCK) Using a Novel Canine DNA Microarray

Daniel F. Balkovetz1, Edward R. Gerrard Jr.2, Shixiong Li1, David Johnson3, James Lee3, John W. Tobias4, Katherine K. Rogers3, Richard W. Snyder3, and Joshua H. Lipschutz5*

1 Department of Medicine and Cell Biology, University of Alabama at Birmingham and Veterans Administration Medical Center Birmingham, Birmingham, AL, USA
2 Department of Surgery, University of Alabama at Birmingham, Birmingham, AL, USA
3 Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA
4 Genomics Institute, Bioinformatics Core, University of Pennsylvania, Philadelphia, PA, USA
5 Depatment of Medicine & Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, PA, USA

* To whom correspondence should be addressed. E-mail: jhlipsch{at}mail.med.upenn.edu.

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, 8 genes previously implicated in the HGF signaling pathway were differentially regulated, demonstrating that the system was responsive to HGF. 117 genes not previously known to be involved in the HGF pathway were also identified. The results were confirmed by real-time PCR or Western blot for 38 genes. Of particular interest were the large number of differentially regulated genes encoding small GTPases, proteins involved in ER translation, proteins involved in the cytoskeleton, the extracellular matrix, and the hematopoietic and prostaglandin systems.




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