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RESIDUAL OIL FLY ASH (ROFA) AND VANADIUM-INDUCED GENE EXPRESSION PROFILES IN HUMAN VASCULAR ENDOTHELIAL CELLS
Citation:
Nadadur, S. S., U P. Kodavanti, MJK Selgrade, AND D L. Costa. RESIDUAL OIL FLY ASH (ROFA) AND VANADIUM-INDUCED GENE EXPRESSION PROFILES IN HUMAN VASCULAR ENDOTHELIAL CELLS. Presented at Lovelace Respiratory Research Institute Annual Symp, Albuquerque, NM, October 13-15, 2002.
Description:
Residual oil fly ash (ROFA) and vanadium-induced gene expression profiles in human vascular endothelial cells.
Srikanth S. Nadadur, Urmila P. Kodavanti, Mary Jane Selgrade and Daniel L. Costa, Pulmonary Toxicology Branch, ETD, NHEERL, ORD, US EPA, Research Triangle Park, NC 27711.
Epidemiological studies have reported a positive association between short-term increases in ambient particulate matter (PM) and increased hospital admission and daily mortality for cardiovascular disease. Whether the observed cardiovascular effects of PM and/or its constituents have direct or indirect impact on the cardiovascular system is unclear. Endothelial cells, that line the vascular tree, serve as a selective thrombogenic barrier between blood components and tissues. Dysfunction of vascular permeability contributes to the pathogenesis of a wide range of diseases. Primary vascular endothelial cell culture models are being utilized to investigate and understand the possible role of endothelial dysfunction in PM cardiovascular toxicity. Acute injury to a cell on exposure to a toxicant can initiate a complex series of biological responses, which in turn link to transcriptional activation or inactivation of genes. Assessing the temporal and functional relationship of differentially expressed genes with global gene expression profiling approaches could reveal novel interactions in the initiation and progression of acute injury. In this study primary cultures of human umbilical vein endothelial cells (HUVEC), (VEC Technologies, Inc., New York, NY) were exposed to saline, ROFA (1mg/ml) or vanadium (V) (1mM) for 20 minutes and the gene expression was analyzed using the human plastic microarray (Clontech, Palo Alto, CA) containing 8,448 sequence verified genes available in the Unigene database. PolyA+ RNA derived from 15mg of total RNA was utilized to prepare 33P-labeled cDNA probe and hybridized to plastic arrays. The gene expression data analyzed using GeneSpring software (Silicon Genetics, Redwood City, CA), revealed exposure-specific differential expression in ~1200 genes. Around 700 genes were induced 5-20 fold and ~500 genes were suppressed (5-10 fold) due to ROFA or V exposure. Further analysis indicated ROFA-, and V- specific differential regulation and a set of genes regulated by both ROFA and V. Differential expression was observed in cytokines, growth factors, cell proliferation, transcription factors, signaling molecules, extra cellular matrix repair genes, transcriptional activation and nuclear regulation pathways. Among the 39 interleukins analyzed, ten were found induced and 19 were found suppressed on exposure to ROFA or V. While induction of IL-14 and interferon-a 10 appear to be specific to ROFA, IL-12b, IL-19 and interferon-b1 were specific to V. Four interleukins IL-13, IL-17, IL-17b and interferon-a14 were induced by both ROFA and V. Hierarchical clustering analysis on the gene expression data generated three unique gene clusters representing exposure specific alteration in gene expression. Differential gene expression data derived here suggests ROFA component specific effects and the possible involvement of endothelial cells in PM toxicity. (This abstract does not reflect US EPA policy).