Effect of aqueous tobacco smoke extract and shear stress on PECAM-1 expression and cell motility in human uterine endothelial cells.
Tobacco smoke constituents have several adverse effects on endothelial cells. Exposure to tobacco smoke during pregnancy is associated with adverse effects on pregnancy outcome possibly related to endothelial dysfunction. Platelet endothelial cell adhesion molecule-1 (PECAM-1) is an important regulator of endothelial function. This study tests the idea that an aqueous extract of cigarette smoke alters the expression of PECAM-1 in uterine endothelial cells. Human uterine microvascular endothelial cells were cultured in cigarette smoke-conditioned medium (CSM) under arterial physiological flow conditions (shear or frictional stress in the range 7.5-15 dyne/cm(2)) and the expression of PECAM-1 was assessed by immunofluorescence microscopy and Western blotting. Thick reticular PECAM-1-associated bands found at cell-cell junctions in static cultures became significantly thinner or disappeared when the cells were exposed to shear stress or to CSM for 24 h. This diminution at cell junctions was accompanied by increased punctate cytoplasmic/cell surface staining. Under shear stress conditions, PECAM-1 was equally distributed between cell surface and intracellular sites. In contrast, when cells were exposed to both shear stress and CSM, PECAM-1 was predominantly localized to the cell surface. It was shown that shear stress increased endothelial cell migration and that CSM abrogated this effect. These results suggest that, under shear stress conditions, PECAM-1 is not predominantly concentrated at intercellular junctions in uterine endothelial cells. Exposure of cells to unidentified soluble components of cigarette smoke leads to alterations in PECAM-1 distribution that may cause endothelial dysfunction. If this occurs in vivo it could contribute to the adverse effects on pregnancy outcome associated with exposure to cigarette smoke.
Soghomonians
Arlen
A
Department of Aeronautical and Mechanical Engineering, University of California, Davis, California 95616, USA.
Thirkill
Twanda L
TL
Mariano
Natalie F
NF
Barakat
Abdul I
AI
Douglas
Gordon C
GC
D015153:Blotting, Western; D002196:Capillaries / Q000166:cytology; D002465:Cell Movement / Q000187:drug effects*; D003470:Culture Media; D003864:Depression, Chemical; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D000432:Methanol; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis* / Q000235:genetics; D011108:Polymers; D011247:Pregnancy; D012906:Smoke / Q000009:adverse effects*; D012997:Solvents; D013314:Stress, Mechanical; D014404:Tubulin / Q000096:biosynthesis / Q000235:genetics; D014599:Uterus / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*
D003470:Culture Media; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011108:Polymers; D012906:Smoke; D012997:Solvents; D014404:Tubulin; D005557:Formaldehyde; C003043:paraform; D000432:Methanol