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NORMAL NASAL GENE EXPRESSION LEVELS USING CDNA ARRAY TECHNOLOGY
Hester, S D., G. R. Benavides, M. Sartor, L. Yoon, K. T. Morgan, AND D C. Wolf. NORMAL NASAL GENE EXPRESSION LEVELS USING CDNA ARRAY TECHNOLOGY. Presented at Annual AACR Mtg, New Orleans, Louisanna, March 19-25, 2001.
Normal Nasal Gene Expression Levels Using cDNA Array Technology.
The nasal epithelium is a target site for chemically-induced toxicity and carcinogenicity. To detect and analyze genetic events which contribute to nasal tumor development, we first defined the gene expression profile in control rats. Genes regulating mucous production, detoxification of xenobiotics, metabolism, and cell turnover are critical to nasal epithelial cell homeostasis. The differential hybridization to cDNA arrays can provide a means to define genetic patterns which underlie and support the normal physiology of cells lining the nasal passages. We first developed a method for the isolation of epithelial cells from this site. Four rats were euthanized, and the head removed. Cells lining the anterior nasal airway passages, as confirmed by light microscopy, were collected into 200 ul of TrizolTM reagent by controlled nasal instillation. Total RNA was used to perform the cDNA array analysis using ClontechTM Rat Atlas 1.2. and verified using real-time PCR, (TaqManTM). The percentages of genes within a specific average expression ranges, based on phosphorimager signal intensities, were 1.4% at 3000-45,000; 3.3% at 1000-3000; 30.1% at 100-1000; 65.2% below 100. The most highly expressed genes included those involved in phase I (e.g. CYPs) and phase II (e.g. GSTs) xenobiotic metabolism, bioenergetics (e.g. cytochrome oxidase), osmotic balance (e.g. Na+/K+ ATPase) and epithelial ionic homeostasis (e.g. ion channels). These data allow further understanding of the biology of normal nasal epithelial gene expression in the rat and enable detection of true alterations of gene expression for defining biologically relevant responses to inhaled toxicants and carcinogens.
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