Effect of L-homocysteine on endothelial cell-cell junctions following F-actin stabilization through tropomyosin-1 overexpression

  • Authors:
    • Maciej Gagat
    • Dariusz Grzanka
    • Magdalena Izdebska
    • Alina Grzanka
  • View Affiliations

  • Published online on: April 22, 2013     https://doi.org/10.3892/ijmm.2013.1357
  • Pages: 115-129
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Abstract

Since the identification of actin in non‑muscle cells, it has been suggested that the regulation of the mechanical behaviors of the actin cytoskeleton regulates cellular shape changes and the generation of forces during cell migration and division. The maintenance of cell shape and polarity are important in the formation of cell-cell junctions. The aim of the present study was to determine the effect of L‑homocysteine thiolactone hydrochloride on EA.hy926 endothelial cells in the context of the maintenance cell-cell junctions through the stabilization of filamentous actin cytoskeleton (F‑actin). The actin filaments were stabilized by the overexpression of tropomyosin-1, which has the ability to stabilize actin filaments in muscle and non-muscle cells. The stabilization of F-actin induced a significant decrease in the percentage of late apoptotic and necrotic cells following treatment with L-homocysteine. Moreover, the migratory potential of the endothelial cells was greater in the cells overexpressing tropomyosin-1 treated with L-homocysteine. Additionally, our results indicated that the stabilization of F-actin in the EA.hy926 cells significantly increased the expression of junctional β‑catenin, as compared to the cells not overexpressing tropomyosin‑1. Similarly, the fluorescence intensity of junctional α-catenin was also increased in the cells with stabilized F‑actin cytoskeleton. However, this increase was only slightly higher than that observed in the EA.hy926 cells not overexpressing tropomyosin-1. Furthermore, the analysis of Zonula occludens (ZO)‑1 relative fluorescence demonstrated a statistically significant decrease in the cell-cell junction areas among the cells with stabilized F-actin cytoskeleton in comparison to the cells not overexpressing tropomyosin-1. Our results indicate that the stabilization of F-actin does not affect the migratory potential of cells, and consequently protects the EA.hy926 cells against the L-homocysteine-induced decrease in cell mobility. Moreover, it is suggested that α‑catenin may participate in the suppression of actin polymerization in the area of cell-cell junctions. It can be hypothesized that the stabilization of F-actin strengthens endothelial adherens and tight junctions by increasing the number of cell-cell junctions due to the amplification of β-catenin and the ZO‑1 fluorescence signal. However, ZO-1 stabilizes the endothelial barrier function through the stabilization of F-actin and F-actin itself stabilizes the localization of ZO-1.
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July 2013
Volume 32 Issue 1

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Gagat M, Grzanka D, Izdebska M and Grzanka A: Effect of L-homocysteine on endothelial cell-cell junctions following F-actin stabilization through tropomyosin-1 overexpression. Int J Mol Med 32: 115-129, 2013
APA
Gagat, M., Grzanka, D., Izdebska, M., & Grzanka, A. (2013). Effect of L-homocysteine on endothelial cell-cell junctions following F-actin stabilization through tropomyosin-1 overexpression. International Journal of Molecular Medicine, 32, 115-129. https://doi.org/10.3892/ijmm.2013.1357
MLA
Gagat, M., Grzanka, D., Izdebska, M., Grzanka, A."Effect of L-homocysteine on endothelial cell-cell junctions following F-actin stabilization through tropomyosin-1 overexpression". International Journal of Molecular Medicine 32.1 (2013): 115-129.
Chicago
Gagat, M., Grzanka, D., Izdebska, M., Grzanka, A."Effect of L-homocysteine on endothelial cell-cell junctions following F-actin stabilization through tropomyosin-1 overexpression". International Journal of Molecular Medicine 32, no. 1 (2013): 115-129. https://doi.org/10.3892/ijmm.2013.1357