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Inflammation Research

, Volume 50, Issue 4, pp 220–226 | Cite as

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

  • Y. Yu
  • C. De Waele
  • K. Chadee

Abstract:

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.¶

Key words: Chemokine - Interleukin-8 - Calcium - Human colonic epithelial cells - Gene expression 

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Copyright information

© Birkhäuser Verlag, 2001

Authors and Affiliations

  • Y. Yu
    • 1
  • C. De Waele
    • 1
  • K. Chadee
    • 1
  1. 1.Institute of Parasitology of McGill University, 21,111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, Canada H9X 3V9, Fax: 514-398-7857 e-mail: kris_chadee@maclan.mcgill.ca CA

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