Gene expression changes associated with cytotoxicity identified using cDNA arrays

M. A. Gore, M. M. Morshedi, J. F. Reidhaar-Olson

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

In order to investigate gene expression changes associated with cytotoxicity, we used cDNA arrays to monitor the expression of over 5,000 genes in response to toxic stress in the HepG2 liver cell line. Cells were treated with cytotoxic doses of acetaminophen, caffeine or thioacetamide for nine time points ranging from 1 to 24 h. Samples of mRNA from each time point were used to prepare radiolabeled cDNA, which was hybridized to nylon-membrane-based cDNA arrays. High-stringency washes were applied to reduce cross-hybridization. Analysis of spot intensities revealed that each compound led to approximately 150-250 gene expression changes that were sustained over at least three adjacent time points. The affected genes could be classified into clusters based on their temporal patterns of differential expression. A common set of 44 genes showed similar expression changes in response to all three compounds. Of these changes, 90% could be confirmed by quantitative RT-PCR analysis. The results indicate that detailed array-based time-course studies, coupled with a sensitive and highly specific confirmation assay, provide a powerful means of identifying cytotoxicity-associated gene expression changes.

Original languageEnglish (US)
Pages (from-to)114-126
Number of pages13
JournalFunctional and Integrative Genomics
Volume1
Issue number2
DOIs
StatePublished - Dec 1 2000

Keywords

  • Cytotoxicity
  • Differential gene expression
  • Expression profiling
  • cDNA arrays

ASJC Scopus subject areas

  • Genetics

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