Calcium-dependent interleukin-8 gene expression in T84 human colonic epithelial cells

Inflamm Res. 2001 Apr;50(4):220-6. doi: 10.1007/s000110050747.


Objective and design: IL-8 is a chemokine that activates and recruits neutrophils and plays a major role in intestinal inflammation. Signal transduction pathways mediated by protein kinases are central in regulating IL-8 gene expression, however, little is known about the role of Ca2+ in this event. In this study, we characterize the effect of intracellular Ca2+ on interleukin-8 gene expression in T84 human colonic epithelial cells.

Materials and methods: Cells were stimulated with Ca2+ ionophore, A23187 or thapsigargin, a Ca2+-ATPase inhibitor. Semi-quantitative RT-PCR was used to examine IL-8 mRNA and ELISA for protein quantification. Reporter gene techniques were used to determine transcription rate.

Results: A23187 and thapsigargin caused a dose- and time-dependent accumulation of IL-8 mRNA and protein production which was dependent on the release of Ca2+ from intracellular stores. FK506, a specific inhibitor of calcineurin, inhibited A23187- and thapsigargin-induced IL-8 mRNA expression in a dose dependent manner. Reporter gene studies and actinomycin D chase experiments showed that A23187 and thapsigargin enhanced IL-8 gene transcription and stabilized IL-8 mRNA transcripts, respectively.

Conclusion: Intracellular Ca2+ plays an important role in regulating IL-8 transcriptionally and posttranscriptionally through calcium/calmodulin-dependent calcineurin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma
  • Calcimycin / pharmacology
  • Calcineurin / pharmacology
  • Calcium / pharmacology*
  • Colon / metabolism*
  • Colonic Neoplasms
  • Dose-Response Relationship, Drug
  • Epithelial Cells / metabolism
  • Gene Expression* / drug effects
  • Humans
  • Interleukin-8 / genetics*
  • Ionophores / pharmacology
  • Kinetics
  • Protein Kinase C / metabolism
  • RNA, Messenger / analysis
  • Tetradecanoylphorbol Acetate / pharmacology
  • Thapsigargin / pharmacology
  • Transcription, Genetic / drug effects
  • Tumor Cells, Cultured


  • Interleukin-8
  • Ionophores
  • RNA, Messenger
  • Calcimycin
  • Thapsigargin
  • Protein Kinase C
  • Calcineurin
  • Tetradecanoylphorbol Acetate
  • Calcium