Cholera toxin-induced modulation of gene expression: elucidation via cDNA microarray for rational cell-based sensor design

被引:13
作者
Lin, HJ
Charles, PT
Andreadis, JD
Churilla, AM
Stenger, DA
Pancrazio, JJ
机构
[1] USN, Res Lab, Ctr Biomol Sci & Engn, Washington, DC 20375 USA
[2] USN, Res Lab, Div Chem, Washington, DC 20375 USA
关键词
cDNA microarray; false positive; cholera; epithelial; gene expression; enterotoxin; lymphoma;
D O I
10.1016/S0003-2670(01)01353-8
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Cell-based biosensors utilize functional changes in cellular response to identify the biological threats in a physiological relevant manner. Cell-based sensors have been used for a wide array of applications including toxicological assessment and drug-screening. In this paper, we utilize DNA arrays to identify differential gene expression events induced by toxin exposure for the purpose of developing a reporter gene assay system compatible with insertion into a cell-based sensor platform. HT29, an intestine epithelial cell line, was used as a cell model to study the cholera toxin (CT)-induced host cell modulation using DNA array analysis. A false positive model was generated from analysis of housekeeping genes in untreated control experiments to characterize our system and to minimize the number of false positives in the data. Threshold probability scores (-3.72), which gives <0.02% false positives for up/down regulation from the false positive model, were used to identify 73 and 25 known genes/expression tag sequences (ESTs) that were up- and down-regulated, respectively, in cells exposed 23 nM of CT. Using quantitative multiplex PCR assay, the gene expression levels for several genes shown to be modulated according to the microarray experiments, such as apolipoprotein D (Apol D), E-cadherin, and cyclin A2, were. confirmed. The differential expression of genes encoding cytochrome P450, glutathione transferase (GST), and MGAT2 were noteworthy and consistent with previous studies. Our study provides an approach to analyze cDNA microarray data with defined false positive rates. The utility of cDNA microarray information for the design of cell-based sensor using a reporter gene approach is discussed. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:97 / 108
页数:12
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