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Molecular and Cellular Biochemistry

, Volume 362, Issue 1–2, pp 71–85 | Cite as

Claudin-5 participates in the regulation of endothelial cell motility

  • Astrid Escudero-Esparza
  • Wen G. Jiang
  • Tracey A. MartinEmail author
Article

Abstract

A key step in metastasis is the interaction and penetration of the vascular endothelium by cancer cells. Tight Junctions (TJ) are located between the cancer epithelial cells and between the endothelial cells functioning in an adhesive manner. They represent a critical barrier which the cancer cells must overcome in order to penetrate and initiate metastasis. Claudin-5 is a protein member of the Claudin family, a group of TJ proteins expressed in both endothelial and epithelial cells. This study examined in vitro the effect of altering levels of expression of Claudin-5 in HECV cells. Insertion of Claudin-5 gene in HECV cells resulted in cells that were significantly less motile and less adhesive to matrix (P < 0.001). These cells also exhibited a significant decreased in the angiogenic potential (P < 0.001). Results also revealed a link between Claudin-5 and cell motility. Furthermore, a possible link between Claudin-5 and N-WASP, and Claudin-5 and ROCK was demonstrated when interactions between these proteins were seen in the cell line. Moreover, followed by treatment of N-WASP inhibitor (Wiskostatin) and ROCK inhibitor (Y-27632), cell motility and angiogenic potential were assessed in response to the inhibitors. Results showed that the knockdown of Claudin-5 in HECV cells masked their response to both N-WASP and ROCK inhibitors. In conclusion, this study portrays a new and interesting role for Claudin-5 in cell motility involving the N-WASP and ROCK signalling cascade which is beyond the primarily role of Claudin-5 in keeping the cell barrier tight as it was originally reported.

Keywords

Claudin-5 Endothelial N-WASP ROCK 

Notes

Acknowledgment

We would like to thank Cancer research Wales for helping to fund this research.

References

  1. 1.
    Cereijido M, Contreras RG, Shoshani L, Flores-Benitez D, Larre I (2008) Tight junction and polarity interaction in the transporting epithelial phenotype. Biochim Biophys Acta 1778(3):770–793PubMedCrossRefGoogle Scholar
  2. 2.
    Martin TA, Das T, Mansel RE, Jiang WG (2006) Synergistic regulation of endothelial tight junctions by antioxidant (Se) and polyunsaturated lipid (GLA) via claudin-5 modulation. J Cell Biochem 98(5):1308–1319PubMedCrossRefGoogle Scholar
  3. 3.
    Paschoud S, Bongiovanni M, Pache JC, Citi S (2007) Claudin-1 and claudin-5 expression patterns differentiate lung squamous cell carcinomas from adenocarcinomas. Mod Pathol 20(9):947–954PubMedCrossRefGoogle Scholar
  4. 4.
    Turunen M, Talvensaari-Mattila A, Soini Y, Santala M (2009) Claudin-5 overexpression correlates with aggressive behavior in serous ovarian adenocarcinoma. Anticancer Res 29(12):5185–5189PubMedGoogle Scholar
  5. 5.
    Tsukita S, Furuse M (2000) Pores in the wall: claudins constitute tight junction strands containing aqueous pores. J Cell Biol 149(1):13–16PubMedCrossRefGoogle Scholar
  6. 6.
    Ohkubo T, Ozawa M (2004) The transcription factor Snail downregulates the tight junction components independently of E-cadherin downregulation. J Cell Sci 117(Pt 9):1675–1685PubMedCrossRefGoogle Scholar
  7. 7.
    Morita K, Furuse M, Fujimoto K, Tsukita S (1999) Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands. Proc Natl Acad Sci USA 96(2):511–516PubMedCrossRefGoogle Scholar
  8. 8.
    Furuse M, Sasaki H, Tsukita S (1999) Manner of interaction of heterogeneous claudin species within and between tight junction strands. J Cell Biol 147(4):891–903PubMedCrossRefGoogle Scholar
  9. 9.
    Tsukita S (1989) Isolation of cell-to-cell adherens junctions from rat liver. J Cell Biol 108(1):31–41PubMedCrossRefGoogle Scholar
  10. 10.
    Van Itallie CM, Anderson JM (2006) Claudins and epithelial paracellular transport. Annu Rev Physiol 68:403–429PubMedCrossRefGoogle Scholar
  11. 11.
    Morita K, Sasaki H, Furuse M, Tsukita S (1999) Endothelial claudin: claudin-5/TMVCF constitutes tight junction strands in endothelial cells. J Cell Biol 147(1):185–194PubMedCrossRefGoogle Scholar
  12. 12.
    Rahner C, Mitic LL, Anderson JM (2001) Heterogeneity in expression and subcellular localization of claudins 2, 3, 4, and 5 in the rat liver, pancreas, and gut. Gastroenterology 120(2):411–422PubMedCrossRefGoogle Scholar
  13. 13.
    Amasheh S, Schmidt T, Mahn M, Florian P, Mankertz J, Tavalali S, Gitter AH, Schulzke JD, Fromm M (2005) Contribution of claudin-5 to barrier properties in tight junctions of epithelial cells. Cell Tissue Res 321(1):89–96PubMedCrossRefGoogle Scholar
  14. 14.
    Nitta T, Hata M, Gotoh S, Seo Y, Sasaki H, Hashimoto N, Furuse M, Tsukita S (2003) Size-selective loosening of the blood–brain barrier in claudin-5-deficient mice. J Cell Biol 161(3):653–660PubMedCrossRefGoogle Scholar
  15. 15.
    Wolburg H, Wolburg-Buchholz K, Kraus J, Rascher-Eggstein G, Liebner S, Hamm S, Duffner F, Grote EH, Risau W, Engelhardt B (2003) Localization of claudin-3 in tight junctions of the blood–brain barrier is selectively lost during experimental autoimmune encephalomyelitis and human glioblastoma multiforme. Acta Neuropathol 105(6):586–592PubMedGoogle Scholar
  16. 16.
    Jiang WG, Davies G, Martin TA, Parr C, Watkins G, Mason MD, Mokbel K, Mansel RE (2005) Targeting matrilysin and its impact on tumor growth in vivo: the potential implications in breast cancer therapy. Clin Cancer Res 11(16):6012–6019PubMedCrossRefGoogle Scholar
  17. 17.
    Martin TA, Pereira G, Watkins G, Mansel RE, Jiang WG (2008) N-WASP is a putative tumour suppressor in breast cancer cells, in vitro and in vivo, and is associated with clinical outcome in patients with breast cancer. Clin Exp Metastasis 25(2):97–108PubMedCrossRefGoogle Scholar
  18. 18.
    Hiscox S, Jiang WG (1999) Association of the HGF/SF receptor, c-met, with the cell-surface adhesion molecule, E-cadherin, and catenins in human tumor cells. Biochem Biophys Res Commun 261(2):406–411PubMedCrossRefGoogle Scholar
  19. 19.
    Sanders AJ, Guo X, Mason MD, Jiang WG (2010) IL-17B can impact on endothelial cellular traits linked to tumour angiogenesis. J Oncol 81:73–75Google Scholar
  20. 20.
    Martin TA, Harrison GM, Watkins G, Jiang WG (2008) Claudin-16 reduces the aggressive behavior of human breast cancer cells. J Cell Biochem 105(1):41–52PubMedCrossRefGoogle Scholar
  21. 21.
    Jiang WG, Hiscox S, Hallett MB, Scott C, Horrobin DF, Puntis MC (1995) Inhibition of hepatocyte growth factor-induced motility and in vitro invasion of human colon cancer cells by gamma-linolenic acid. Br J Cancer 71(4):744–752PubMedCrossRefGoogle Scholar
  22. 22.
    Keese CR, Wegener J, Walker SR, Giaever I (2004) Electrical wound-healing assay for cells in vitro. Proc Natl Acad Sci USA 101(6):1554–1559PubMedCrossRefGoogle Scholar
  23. 23.
    Mitchison TJ, Cramer LP (1996) Actin-based cell motility and cell locomotion. Cell 84(3):371–379PubMedCrossRefGoogle Scholar
  24. 24.
    Mitchison TJ, Cramer LP (1996) Loosening of the blood–brain barrier in claudin-5-deficient mice. J Cell Biol 161(3):653–660Google Scholar
  25. 25.
    Short SM, Talbott GA, Juliano RL (1998) Integrin-mediated signaling events in human endothelial cells. Mol Biol Cell 9(8):1969–1980PubMedGoogle Scholar
  26. 26.
    Arnaoutova I, George J, Kleinman HK, Benton G (2009) The endothelial cell tube formation assay on basement membrane turns 20: state of the science and the art. Angiogenesis 12(3):267–274PubMedCrossRefGoogle Scholar
  27. 27.
    Jiang WG, Martin TA, Parr C, Davies G, Matsumoto K, Nakamura T (2005) Hepatocyte growth factor, its receptor, and their potential value in cancer therapies. Crit Rev Oncol Hematol 53(1):35–69PubMedCrossRefGoogle Scholar
  28. 28.
    Dovas A, Cox D (2010) Regulation of WASp by phosphorylation: activation or other functions? Commun Integr Biol 3(2):101–105PubMedCrossRefGoogle Scholar
  29. 29.
    Lane J, Martin TA, Watkins G, Mansel RE, Jiang WG (2008) The expression and prognostic value of ROCK I and ROCK II and their role in human breast cancer. Int J Oncol 33(3):585–593PubMedGoogle Scholar
  30. 30.
    Osiak AE, Zenner G, Linder S (2005) Subconfluent endothelial cells form podosomes downstream of cytokine and RhoGTPase signaling. Exp Cell Res 307(2):342–353PubMedCrossRefGoogle Scholar
  31. 31.
    Li B, Zhao WD, Tan ZM, Fang WG, Zhu L, Chen YH (2006) Involvement of Rho/ROCK signalling in small cell lung cancer migration through human brain microvascular endothelial cells. FEBS Lett 580(17):4252–4260PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Astrid Escudero-Esparza
    • 1
  • Wen G. Jiang
    • 1
  • Tracey A. Martin
    • 1
    Email author
  1. 1.Metastasis and Angiogenesis Research Group, University Department of SurgeryCardiff University School of MedicineCardiffUK

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