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TNFα up-regulates claudin-2 expression in epithelial HT-29/B6 cells via phosphatidylinositol-3-kinase signaling

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Abstract

Our aim has been to characterize the molecular mechanisms regulating the expression of the channel-forming tight-junctional protein claudin-2 in response to the pro-inflammatory cytokine tumor necrosis factor-α (TNFα), which is elevated, for example, in active Crohn’s disease. TNFα caused an 89% decrease of the paracellular resistance in colonic HT-29/B6 cells, whereas transcellular resistance was unaltered. The claudin-2 protein level was increased by TNFα without changes in subcellular tight-junctional protein localization as revealed by confocal laser scanning microscopy. Enhanced gene expression was identified as the source of this increase, since claudin-2-specific mRNA and promoter activity was elevated, whereas mRNA stability remained unaltered. Specific inhibitors and phospho-specific antibodies revealed that the increased gene expression of claudin-2 after TNFα treatment was mediated by the phosphatidylinositol-3-kinase pathway. Thus, the up-regulation of claudin-2 by TNFα is attributable to the regulation of the expression of the gene, as a result of which epithelial barrier function is disturbed, for example, during chronic intestinal inflammation.

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Acknowledgements

We thank Nicole Held and Susanna Schön for excellent technical assistance. The support of Detlef Sorgenfrei (electronic engineer) is gratefully acknowledged.

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Correspondence to J. D. Schulzke.

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J. Mankertz and M. Amasheh contributed equally to this work.

This work was supported by the Deutsche Forschungsgemeinschaft and the Sonnenfeld-Stiftung Berlin.

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Mankertz, J., Amasheh, M., Krug, S.M. et al. TNFα up-regulates claudin-2 expression in epithelial HT-29/B6 cells via phosphatidylinositol-3-kinase signaling. Cell Tissue Res 336, 67–77 (2009). https://doi.org/10.1007/s00441-009-0751-8

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