Gene expression alterations during HGF-induced dedifferentiation of a renal tubular epithelial cell line (MDCK) using a novel canine DNA microarray.
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.
Balkovetz
Daniel F
DF
Departments of Medicine and Cell Biology, University of Alabama at Birmingham, and Veterans Administration Medical Center, Birmingham, Alabama 35294, USA.
Gerrard
Edward R
ER
Li
Shixiong
S
Johnson
David
D
Lee
James
J
Tobias
John W
JW
Rogers
Katherine K
KK
Snyder
Richard W
RW
Lipschutz
Joshua H
JH
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D004285:Dogs; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D007684:Kidney Tubules / Q000166:cytology / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*
D017228:Hepatocyte Growth Factor