Analysis of interleukin-1beta-modulated mRNA Gene Transcription in Human Gingival Keratinocytes by Epithelia-Specific cDNA Microarrays

J Periodontal Res. 2006 Oct;41(5):426-46. doi: 10.1111/j.1600-0765.2006.00884.x.

Abstract

Background/objectives: Proinflammatory cytokines such as interleukin-1beta are known to be synthesized in oral gingivitis and periodontitis and lead to the activation of the transcription factor nuclear factor-kappaB (NF-kappaB). Although numerous effects of interleukin-1beta on mesenchymal cells are known, e.g. up-regulation of intercellular adhesion molecule-1 in endothelial cells, little is known of the effects of interleukin-1beta on oral keratinocytes. The purpose of the present study was to seek interleukin-1beta-mediated alterations in mRNA gene transcription and a putative activation of NF-kappaB in oral gingival keratinocytes.

Methods: As an in vitro model for gingivitis and periodontitis, immortalized human gingival keratinocytes (IHGK) were stimulated with the proinflammatory cytokine interleukin-1beta. An epithelia-specific cDNA microarray was used to analyze mRNA expression profiles from IHGK cells treated with 200 units interleukin-1beta/ml for 3, 6, 9, 12, and 24 h. Indirect immunofluorescence was carried out to detect NF-kappaB in IHGK following interleukin-1beta treatment.

Results: Detailed analysis revealed distinct patterns of time-dependent changes, including genes induced or repressed early (3-6 h) or late (12-24 h) after interleukin-1beta treatment. Differentially expressed genes were involved in (i) cell stress, (ii) DNA repair, (iii) cell cycle and proliferation, (iv) anti-pathogen response, (v) extracellular matrix turnover, and (vi) angiogenesis. A large number of genes were responsive to NF-kappaB and induction was concomitant with nuclear translocation of the p65 RelA subunit of NF-kappaB. Interestingly, many of these genes contain multiple NF-kappaB binding sites in their promoters.

Conclusion: Analysis of altered gene expression allows identification of gene networks associated with inflammatory responses. In addition to a number of well-known genes involved in gingivitis and periodontitis, we identified novel candidates that might be associated with the onset and maintenance of an inflammatory disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / biosynthesis
  • Cell Line, Transformed
  • Cytokines / biosynthesis
  • DNA Repair Enzymes / biosynthesis
  • Extracellular Matrix Proteins / biosynthesis
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression Profiling
  • Gene Regulatory Networks
  • Gingiva / cytology
  • Gingiva / metabolism*
  • Gingivitis / genetics*
  • Gingivitis / metabolism
  • Heat-Shock Proteins / biosynthesis
  • Humans
  • Interleukin-1beta / physiology*
  • Keratinocytes / metabolism*
  • Matrix Metalloproteinases / biosynthesis
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Oligonucleotide Array Sequence Analysis / methods
  • Promoter Regions, Genetic
  • Protein Transport
  • RNA, Messenger / biosynthesis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcriptional Activation / physiology*
  • Vascular Endothelial Growth Factor A / biosynthesis

Substances

  • Cell Cycle Proteins
  • Cytokines
  • Extracellular Matrix Proteins
  • Heat-Shock Proteins
  • Interleukin-1beta
  • NF-kappa B
  • RNA, Messenger
  • Vascular Endothelial Growth Factor A
  • Matrix Metalloproteinases
  • DNA Repair Enzymes