Development of high-density DNA microarray membrane for profiling smoke- and hydrogen peroxide-induced genes in a human bronchial epithelial cell line.
Development of the high-density DNA microarray technique permits the analysis of thousands of genes simultaneously for their differential expression patterns in various biological processes. Through clustering analysis and pattern recognition, the significance of differentially expressed genes can be recognized and correlated with biological events that may take place inside the cell and tissue. With this notion in mind, high-density DNA microarray nylon membrane with colorimetry detection was used to profile the expression of smoke- and hydrogen peroxide-inducible genes in a human bronchial epithelial cell line, HBE1. On the basis of the time course of expression, at least three phases of change in gene expression could be recognized. The first phase is an immediate event in response to oxidant injury. This phase includes induction of the bcl-2 and mdm-2 genes, which are involved in the regulation of apoptosis, and the mitogen-activated protein (MAP) kinase phosphatase 1 (MKP-1) gene, that functions as a regulator of various mitogen-activated protein kinase activities. The second phase, usually 5 h later, includes the induction of various stress proteins and ubiquitin, which are important in providing the chaperone mechanism and the turnover of damaged macromolecules. The third phase, which is 5-10 h later, includes the induction of genes that are apparently involved in reducing oxidative stress by metabolizing reactive oxygen species. In this phase, enzymes associated with tissue and cell remodeling are also elevated. These results demonstrate a complex gene expression array by bronchial epithelial cells in response to the insult of oxidants that are relevant to environmental pollutants.
Yoneda
K
K
Center for Comparative Respiratory Biology and Medicine, Department of Internal Medicine, University of California at Davis, California 95616, USA.
Peck
K
K
Chang
M M
MM
Chmiel
K
K
Sher
Y P
YP
Chen
J
J
Yang
P C
PC
Chen
Y
Y
Wu
R
R
D017209:Apoptosis; D015152:Blotting, Northern; D001980:Bronchi / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D002151:Calorimetry; D002460:Cell Line; D004247:DNA / Q000032:analysis; D018076:DNA, Complementary / Q000032:analysis; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D016877:Oxidants / Q000494:pharmacology*; D018384:Oxidative Stress / Q000235:genetics*; D012333:RNA, Messenger / Q000302:isolation & purification; D017382:Reactive Oxygen Species / Q000378:metabolism; D013997:Time Factors; D014028:Tobacco Smoke Pollution / Q000009:adverse effects*; D025801:Ubiquitin
D018076:DNA, Complementary; D006360:Heat-Shock Proteins; D016877:Oxidants; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D014028:Tobacco Smoke Pollution; D025801:Ubiquitin; D004247:DNA; D006861:Hydrogen Peroxide