Effects of An E-cadherin-Derived Peptide on the Gene Expression of Caco-2 Cells
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Purpose. The goal of this study was to determine the effects of exposure to an HAV peptide (Ac-SHAVSS-NH2) on the protein and gene expression in Caco-2 cells, a model for the intestinal mucosa.
Methods. Caco-2 cells were incubated with either 100 or 500 μM of the hexapeptide then evaluated over a 48-h time period.
Results. Cell detachment from the monolayer was seen only after 48 h of exposure to the peptide, with the greatest effects occurring with a peptide concentration of 500 μM. Total protein expression of E-cadherin showed a decrease of nearly 20% at the 24-h time point for each concentration examined, whereas no significant changes were detected at the other time points studied. Short term exposure to a 500 μM solution of Ac-SHAVSS-NH2 caused few changes in gene expression as determined by Affymetrix GeneChip⌖ microarrays; however, longer exposure periods produced numerous changes in the treated cells. The variations in mRNA expression indicate that this HAV peptide has an effect in the E-cadherin signaling pathways. The greatest increases in mRNA expression were found in genes regulating excretion or degradation of the peptide.
Conclusions. This work suggests that this HAV peptide produces effects that reach beyond modulation of adhesion.
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- 7.K. L. Lutz and T. J. Siahaan. Modulation of the cellular junctions protein E-cadherin in bovine brain microvessel endothelial cells by cadherin peptides. Drug Deliv. 10:187–193 (1997).Google Scholar
- 12.I. Vietor, T. Bader, K. Paiha, and L. A. Huber. Perturbation of the tight junction permeability barrier by occludin loop peptides activates ta-catenin/TCF/LEF-mediated transcription. EMBO Rep. 2:306–312 (2001).Google Scholar
- 17.M. R. Bootcov, A. R. Bauskin, S. M. Valenzuela, A. G. Moore, M. Bansal, X. Y. He, H. P. Zhang, M. Donnellan, S. Mahler, K. Pryor, B. J. Walsh, R. C. Nicholson, W. D. Fairlie, S. B. Por, J. M. Robbins, and S. N. Breit. MIC-1, a novel macrophage inhibitory cytokine, is a divergent member of the TGF-beta superfamily. Proc. Natl. Acad. Sci. USA 94:11514–11519 (1997).PubMedGoogle Scholar
- 18.P. X. Li, J. Wong, A. Ayed, D. Ngo, A. M. Brade, C. Arrowsmith, R. C. Austin, and H. J. Klamut. Placental transforming growth factor-beta is a downstream mediator of the growth arrest and apoptotic response of tumor cells to DNA damage and p53 over-expression. J. Biol. Chem. 275:20127–20135 (2000).PubMedGoogle Scholar
- 19.K. Melen, P. Keskinen, T. Ronni, T. Sareneva, K. Lounatmaa, and I. Julkunen. Human MxB protein, an interferon-alpha-inducible GTPase, contains a nuclear targeting signal and is localized in the heterochromatin region beneath the nuclear envelope. J. Biol. Chem. 271:23478–23486 (1996).PubMedGoogle Scholar
- 21.S. Saunier, J. Calado, R. Heilig, F. Silbermann, F. Benessy, G. Morin, M. Konrad, M. Broyer, M. C. Gubler, J. Weissenbach, and C. Antignac. A novel gene that encodes a protein with a putative src homology 3 domain is a candidate gene for familial juvenile nephronophthisis. Hum. Mol. Genet. 6:2317–2323 (1997).PubMedGoogle Scholar
- 29.T. Kayano, C. F. Burant, H. Fukumoto, G. W. Gould, Y. S. Fan, R. L. Eddy, M. G. Byers, T. B. Shows, S. Seino, and G. I. Bell. Human facilitative glucose transporters. Isolation, functional characterization, and gene localization of cDNAs encoding an isoform (GLUT5) expressed in small intestine, kidney, muscle, and adipose tissue and an unusual glucose transporter pseudo-gene-like sequence (GLUT6). J. Biol. Chem. 265:13276–13282 (1990).PubMedGoogle Scholar
- 32.S. K. Bohm, W. Kong, D. Bromme, S. P. Smeekens, D. C. Anderson, A. Connolly, M. Kahn, N. A. Nelken, S. R. Coughlin, D. G. Payan, and N. W. Bunnett. Molecular cloning, expression and potential functions of the human proteinase-activated receptor-2. Biochem. J. 314:1009–1016 (1996).PubMedGoogle Scholar
- 34.E. M. Kajkowski, C. F. Lo, X. Ning, S. Walker, H. J. Sofia, W. Wang, W. Edris, P. Chanda, E. Wagner, S. Vile, K. Ryan, B. McHendry-Rinde, S. C. Smith, A. Wood, K. J. Rhodes, J. D. Kennedy, J. Bard, J. S. Jacobsen, and B. A. Ozenberger. beta-Amyloid peptide-induced apoptosis regulated by a novel protein containing a g protein activation module. J. Biol. Chem. 276: 18748–18756 (2001).PubMedGoogle Scholar
- 37.E. T. Kipreos and M. Pagano. The F-box protein family. Genome Biol. 1:3002.1-3002.7 (2000).Google Scholar
- 39.S. Wiemann, B. Weil, R. Wellenreuther, J. Gassenhuber, S. Glassl, W. Ansorge, M. Bocher, H. Blocker, S. Bauersachs, H. Blum, J. Lauber, A. Dusterhoft, A. Beyer, K. Kohrer, N. Strack, H. W. Mewes, B. Ottenwalder, B. Obermaier, J. Tampe, D. Heubner, R. Wambutt, B. Korn, M. Klein, and A. Poustka. Toward a catalog of human genes and proteins: sequencing and analysis of 500 novel complete protein coding human cDNAs. Genome Res. 11:422–435 (2001).PubMedGoogle Scholar