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Physiol. Genomics 26: 99-108, 2006. First published April 4, 2006; doi:10.1152/physiolgenomics.00152.2005
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Received 30 June 2005; accepted in final form 22 February 2006.
Physiological Genomics 26:99-108 (2006)
1094-8341/06 $8.00 © 2006 American Physiological Society

Comparative SAGE analysis of the response to hypoxia in human pulmonary and aortic endothelial cells

D. G. Peters 1,2,5,*, W. Ning1,2,3,*, T. J. Chu4, C. J. Li1,2 and A. M. K. Choi1,2

1 Division of Pulmonary, Allergy and Critical Care Medicine, University of Pittsburgh, Pittsburgh
2 Lung Translational Genomics Center, Department of Medicine, University of Pittsburgh, Pittsburgh
3 Model Animal Research Center, Nanjing University, Nanjing, China
4 Institute for Human and Machine Cognition, University of West Florida, Pensacola, Florida
5 Department of Pharmacology and Therapeutics, University of Liverpool, Liverpool, United Kingdom

We utilized serial analysis of gene expression (SAGE) to analyze the temporal response of human pulmonary artery endothelial cells (HPAECs) to short-term chronic hypoxia at the level of transcription. Primary cultures of HPAECs were exposed to 1% O2 hypoxia for 8 and 24 h and compared with identical same-passage cells cultured under standard (5% CO2-95% air) conditions. Hierarchical clustering of significant hypoxia-responsive genes identified temporal changes in the expressions of a number of well-described gene families including those encoding proteins involved in thrombosis, stress response, apoptosis, angiogenesis, and cell proliferation. These experiments build on previously published data describing the transcriptomic response of human aortic endothelial cells (HAECs) obtained from the same donor and cultured under identical conditions, and we have thus taken advantage of the immortality of SAGE data to make direct comparisons between these two data sets. This approach revealed comprehensive information relating to the similarities and differences at the level of mRNA expression between HAECs and HPAECs. For example, we found differences in the cell type-specific response to hypoxia among genes encoding cytoskeletal factors, including paxillin, and proteins involved in metabolic energy production, the response to oxidative stress, and vasoreactivity (e.g., endothelin-1). These efforts contribute to the expanding collection of publicly available SAGE data and provide a foundation on which to base further efforts to understand the characteristics of the vascular response to hypoxia in the pulmonary circulation relative to systemic vasculature.

transcriptome; temporal; serial analysis of gene expression







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