Differential gene expression in young and senescent endothelial cells under static and laminar shear stress conditions

Identification of CD44 as a senescence-induced cell adhesion gene responsible for the enhanced monocyte recruitment to senescent endothelial cells

“…The results of the present study, however, indicated that this would not be the case, because the CD44 expression level was not affected by LSS (Fig. 3), which increased NO production even in senescent endothelial cells (27). Thus the inhibitory effect of LSS and/or NO on leukocyte adhesion, if any, may not be related to CD44 suppression.…”

“…In an approach to identify the genes responsible for enhanced monocyte adhesion to senescent endothelial cells, the cDNA microarray data on the gene expression in young and senescent HUVECs under static and LSS conditions were analyzed (27). Cellular senescence significantly altered the expression levels of 2,351 genes under static conditions and 3,444 genes under LSS conditions.…”

“…Human umbilical vein endothelial cells (HUVECs) obtained from Clonetics Cambrex (Rockland, ME) were cultured in EBM-2 medium containing endothelial growth supplements (Clonetics Cambrex), 10% FBS (GIBCO-BRL, Grand Island, NY), and antibiotics (100 U/ml penicillin, 100 g/ml streptomycin, and 0.25 g/ml amphotericin B) on 0.2% gelatin-coated 100-mm tissue culture dishes (BD Biosciences, San Jose, CA) at 37°C and 5% CO 2 (26). Cells were serially subcultured at ratios of 1 to 3ϳ5 in the same media, until they evidenced senescence-associated phenotypes (27). Cell morphology was evaluated under an Eclipse TS100 inverted phase microscope from Nikon (Melville, NY).…”

“…LSS was provided using a rotating Teflon cone (0.5°Cone angle) mounted on a culture dish, as previously described (2,32). Total cellular RNA was extracted from cells using the RNeasy kit (Qiagen, Valencia, CA), and gene expression profiles were analyzed on a GeneChip HG-U133 Plus 2.0 (Affymetrix, Santa Clara, CA) (27). Three chips were used for each condition.…”

Increased Monocytic Adhesion by Senescence in Human Umbilical Vein Endothelial Cells

“…Senescent vascular endothelial cells have been observed in the atherosclerotic plaques of human coronary arteries 6) which may suggest a relationship between aging of these cells and atherosclerosis. The senescence of endothelial cells is induced by oxidative stress and shear stress [6][7][8] and promotes inflammation and atherosclerosis. 7) However, it is unclear if replicative senescence (i.e., ''pure'' senescence) contributes to the pathogenesis of atherosclerosis.…”