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1 Department of Animal Science and Faculty of Nutrition, Texas A&M University; 2 Cardiovascular Research Institute and Department of Medical Physiology, Texas A&M University System Health Science Center, College Station, Texas, 77843; and 3 Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261
Polyamines are essential for cell proliferation;
therefore, we hypothesized that arginase I or arginase II activities,
via production of ornithine for polyamine synthesis, may be limiting for proliferation of endothelial cells (EC). Bovine coronary venular EC
stably transfected with a lacZ gene (lacZ-EC, control), rat arginase I cDNA (AI-EC), or mouse arginase II cDNA (AII-EC) were utilized to test this hypothesis. Cell-proliferation assays showed that
EC proliferation was markedly increased in AI-EC and AII-EC compared
with lacZ-EC. Expression of proliferating cell nuclear antigen was also
enhanced in AI-EC and AII-EC. DL-
-difluoromethylornithine (DFMO), an
irreversible inhibitor of ornithine decarboxylase, was used to
establish that increased polyamine synthesis was involved in mediating
the enhanced growth of AI-EC and AII-EC. Addition of 5 mM DFMO to the
culture medium completely abolished the differences in cellular
putrescine concentrations and reduced the differences in spermidine
concentrations among AI-EC, AII-EC, and lacZ-EC. The DFMO treatment
also prevented an increase in AI-EC and AII-EC proliferation compared
with lacZ-EC. Addition of 10 and 50 µM putrescine dose-dependently
increased AI-EC, AII-EC, and lacZ-EC growth to the same extent. These
results demonstrate that either arginase isoform can potentially play a
role in modulating EC proliferation by regulating polyamine synthesis.
ornithine; polyamines; cell transfection
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