title: Tropomyosin-1 protects endothelial cell-cell junctions against cigarette smoke extract through F-actin stabilization in EA.hy926 cell line.
abstract: The aim of the study was to estimate the effect of cigarette smoke extract (CSE) on EA.hy926 endothelial cells in culture in the context of maintenance of cell-cell junctions through the structural stabilization of the actin cytoskeleton. In the present study, F-actin was stabilized by the overexpression of tropomyosin-1, which is known to stabilize actin filaments in muscle and non-muscle cells. Our study showed that the stabilization of F-actin significantly increased the survival of cells treated with 25% CSE. In addition, after stabilization of F-actin the migratory potential of EA.hy926 cells subjected to CSE treatment was increased. Our results also showed increased fluorescence intensity of alpha- and beta-catenin after CSE treatment in cells which had stabilized F-actin. Analysis of fluorescence intensity of Zonula occludens-1 did not reveal any significant differences when EA.hy926 cells overexpressing tropomyosin-1 were compared with those lacking overexpression. It would appear that overexpression of tropomyosin-1 preserved the structure of actin filaments in the cells treated with CSE. In conclusion, the present study demonstrates that stabilization of F-actin protects EA.hy926 cells against CSE-induced loss of both adherens and tight junctions. The data presented in this study suggest that overexpression of tropomyosin-1 stabilizes the organizational structure of actin filaments and helps preserve the endothelial barrier function under conditions of strong oxidative stress.
authors.affiliation: Department of Histology and Embryology, Nicolaus Copernicus University in Toruń, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Gagat
authors.affiliation: Department and Clinic of Dermatology, Sexually Transmitted Diseases and Immunodermatology, Nicolaus Copernicus University in Toruń, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
authors.forename: Dariusz
authors.initials: D
authors.lastname: Grzanka
authors.affiliation: Department of Histology and Embryology, Nicolaus Copernicus University in Toruń, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Izdebska
authors.affiliation: Department of Medicinal Chemistry, Nicolaus Copernicus University in Toruń, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
authors.forename: Wiktor Dariusz
authors.initials: WD
authors.lastname: Sroka
authors.affiliation: Department of Medicinal Chemistry, Nicolaus Copernicus University in Toruń, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
authors.forename: Michał Piotr
authors.initials: MP
authors.lastname: Marszałł
authors.affiliation: Department of Histology and Embryology, Nicolaus Copernicus University in Toruń, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland. Electronic address: agrzanka@cm.umk.pl.
authors.forename: Alina
authors.initials: A
authors.lastname: Grzanka
mesh_terms: D000199:Actins / Q000378:metabolism*; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D018384:Oxidative Stress / Q000502:physiology; D019108:Tight Junctions / Q000378:metabolism*; D014026:Nicotiana / Q000737:chemistry*; D014335:Tropomyosin / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism
chemical_list: D000199:Actins; C495270:CTNNB1 protein, human; C490775:TPM1 protein, human; D014335:Tropomyosin; D051176:beta Catenin
keywords: Adherens junctions (AJs); F-actin; Tight junctions (TJs); Tropomyosin-1