Diminished tumor necrosis factor-alpha mediated protein kinase c activity in passaged bovine bronchial epithelial cells

Todd A Wyatt, T. Vcys, S. Emanuel, J. R. Spurzem

Research output: Contribution to journalArticle

Abstract

Epithelial cells play an important role in mediating the effects of pro-inflammatory cytokine.s, such as tumor necrosis facto r-alpha (TNF-a), in the airways. Previously, we have identified that TNF-a stimulates the migration of bovine bronchial epithelial cells (BBEC) in cultured monolayers and one pathway implicated in the transduetion of TNF-a signaling is protein kinase C (PKC). However, the signal transduetion mechanism of bronchial epithelial cell migration is not well understood and the role of protein kinase C (PKC) in these cells has not been extensively investigated. In this study, the subcellular distribution and the enzyme activity of multiple PKC isoforms was investigated in primary vs. passaged cultures of BBEC. We have identified the presence of a, ,. 2, and 6 PKC isoforms in primary cultures of BBEC by Western blot and immunocytochemisiry. PKC activity was demonstrated in subcellular fractions of primary BBEC by assaying Ca-2-dependent PKC activity. TNF-a (5(K) U/ml) induces an increase in migration in primary cultures of BBEC and this response is correlated with the lime of increased PKC activation. Both migration and PKC activity were inhibited by 1 [4M ealphosiin C. Late passage BBEC crude homogenates fractionated by DEAEKophacel Chromatograph y demonstrated the loss of a distinct peak of PKC activity eluting at (O M NaCl, corresponding to ihi? loss of PKC β12, und 8 immunoreuctivity. Additionally, PKC activity in response to TNF-u is decreased in long term cultures of passaged BBEC. We hypoihesi/e that this loss of PKC activity may directly relate to changes in cylokine responsiveness of bronchial epithelial cells of various cellular differentiation states- Supported by Department of Veterans Affairs Research Services.

Original languageEnglish (US)
Pages (from-to)255a
JournalJournal of Investigative Medicine
Volume44
Issue number3
StatePublished - Jan 1 1996

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Protein Kinases
Protein Kinase C
Tumor Necrosis Factor-alpha
Epithelial Cells
Tumors
Necrosis
Neoplasms
Protein Isoforms
Subcellular Fractions
Enzyme activity
Veterans
Cell Movement
Monolayers
Western Blotting
Chemical activation
Cytokines

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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Diminished tumor necrosis factor-alpha mediated protein kinase c activity in passaged bovine bronchial epithelial cells. / Wyatt, Todd A; Vcys, T.; Emanuel, S.; Spurzem, J. R.

In: Journal of Investigative Medicine, Vol. 44, No. 3, 01.01.1996, p. 255a.

Research output: Contribution to journalArticle

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