Stimulation of protein kinase C activity by tumor necrosis factor-α in bovine bronchial epithelial cells

Todd A. Wyatt, Harumasa Ito, Thomas J. Veys, John R. Spurzem

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Abstract

Bronchial epithelial cell migration, attachment, and proliferation are important processes in response to airway injury. We have shown that tumor necrosis factor (TNF)-α stimulates the migration of bovine bronchial epithelial cells (BBEC) in vitro. We hypothesized that protein kinase C (PKC) may be one of the intracellular signaling mediators of TNF-α in BBEC. In this study, we have identified multiple PKC isoforms in BBEC and measured total cellular PKC activity. Polyclonal antibodies to the PKC-α, -β2, - δ, and -ε isoforms recognized protein bands around 80-90 kDa. BBEC primary cultures treated with either 500 U/ml TNF-α for 2-4 h or 100 ng/ml 12-O- tetradecanoylphorbol 13-acetate for 15 min resulted in three- to fivefold increases in PKC activity in the particulate fractions of crude cell lysates. This activity was inhibited by 1 μM calphostin C or 10 μM H-7. Similarly, TNF-α-stimulated BBEC migration was reduced at least twofold in the presence of H-7 or calphostin C. These studies suggest that the activation of PKC is necessary for TNF-α-stimulated BBEC migration.

Original languageEnglish (US)
Pages (from-to)L1007-L1012
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume273
Issue number5 17-5
StatePublished - Dec 9 1997

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Keywords

  • Enzyme activation
  • Epithelial cell migration
  • Protein kinase C isoenzymes

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

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