Genomic and proteomic profiling of oxidative stress response in human diploid fibroblasts

Lifang Xie, Ritu Pandey, Beibei Xu, George Tsaprailis, Qin M. Chen

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A number of lines of evidence suggest that senescence of normal human diploid fibroblasts (HDFs) in culture is relevant to the process of aging in vivo. Using normal human skin diploid fibroblasts, we examine the changes in genes and proteins following treatment with a mild dose of H2O2, which induces premature senescence. Multidimensional Protein Identification Technology (MudPIT) in combination with mass spectrometry analyses of whole cell lysates from HDFs detected 65 proteins in control group, 48 proteins in H2 O2-treated cells and 109 proteins common in both groups. In contrast, cDNA microarray analyses show 173 genes up-regulated and 179 genes down-regulated upon H2O2 treatment. Both MudPIT and cDNA microarray analyses indicate that H2 O2 treatment caused elevated levels of thioredoxin reductase 1. Semi-quantitative RT-PCR and Western-blot were able to verify the finding. Out of a large number of genes or proteins detected, only a small fraction shows the overlap between the outcomes of microarray versus proteomics. The low overlap suggests the importance of considering proteins instead of transcripts when investigating the gene expression profile altered by oxidative stress.

Original languageEnglish (US)
Pages (from-to)125-151
Number of pages27
JournalBiogerontology
Volume10
Issue number2
DOIs
StatePublished - Jan 1 2009

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Keywords

  • Gene expression
  • Human diploid fibroblasts
  • Oxidative stress
  • Proteomics
  • Senescence

ASJC Scopus subject areas

  • Aging
  • Gerontology
  • Geriatrics and Gerontology

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