Effect of endothelial cell proliferation on atherogenesis: A role of p21Sdi/Cip/Waf1 in monocyte adhesion to endothelial cells



      Uniform laminar shear stress (LS) and disturbed turbulent shear stress (DS) are thought to play opposite roles in preventing or inducing atherosclerosis. Endothelial cell (EC) growth and monocyte adhesion to ECs, an early event in atherosclerosis, are also oppositely regulated by LS and DS. However, how atherogenesis is affected by the regulation of growth by blood flow is unknown. Here we examined the role of p21Sdi/Cip/Waf1 (p21), a growth inhibitor induced by LS, in monocyte adhesion to ECs.


      p21 was overexpressed by transfecting a p21-expressing adenoviral vector into ECs. Factors linking EC growth, monocyte adhesion, and p21 were examined by microarray analysis, PCR and Western blotting.


      Compared with DS, in the presence or absence of TNFα, LS significantly inhibited EC growth and monocyte adhesion to ECs. Both EC proliferation and monocyte adhesion induced by DS were inhibited by p21-overexpression. LS suppressed the expression of thioredoxin-interacting protein (TXNIP). Thioredoxin (TRX) activity, which is suppressed by TXNIP, was therefore higher under LS than DS, as reported previously. p21-overexpression significantly suppressed the DS-induced TXNIP expression, and inhibited the expression of vascular cell adhesion molecule 1 and chemokine (C–C motif) ligand 5 (CCL5/RANTES), which stimulates leukocyte recruitment and is downregulated by ROS scavenging.


      p21 may function to prevent atherogenesis by regulating the redox balance, which leads to the inhibition of adhesion molecule and chemokine expression in ECs under LS.


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        • Davies P.F.
        • Remuzzi A.
        • Gordon E.J.
        • Dewey Jr., C.F.
        • Gimbrone Jr., M.A.
        Turbulent fluid shear stress induces vascular endothelial cell turnover in vitro.
        Proc Natl Acad Sci USA. 1986; 83: 2114-2117
        • Levesque M.J.
        • Nerem R.M.
        • Sprague E.A.
        Vascular endothelial cell proliferation in culture and the influence of flow.
        Biomaterials. 1990; 11: 702-707
        • Akimoto S.
        • Mitsumata M.
        • Sasaguri T.
        • Yoshida Y.
        Laminar shear stress inhibits vascular endothelial cell proliferation by inducing cyclin-dependent kinase inhibitor p21Sdi1/Cip1/Waf1.
        Circ Res. 2000; 86: 185-190
        • Chang M.W.
        • Barr E.
        • Lu M.M.
        • Barton K.
        • Leiden J.M.
        Adenovirus-mediated over-expression of the cyclin/cyclin-dependent kinase inhibitor, p21 inhibits vascular smooth muscle cell proliferation and neointima formation in the rat carotid artery model of balloon angioplasty.
        J Clin Invest. 1995; 96: 2260-2268
        • Ueno H.
        • Masuda S.
        • Nishio S.
        • Li J.-J.
        • Yamamoto H.
        • Takeshita A.
        Adenovirus-mediated transfer of cyclin-dependent kinase inhibitor-p21 suppresses neointimal formation in the balloon-injured rat carotid arteries in vivo.
        Ann NY Acad Sci. 1997; 811: 401-411
        • Wara A.K.M.K.
        • Mitsumata M.
        • Yamane T.
        • Kusumi Y.
        • Yoshida Y.
        Gene expression in endothelial cells and intimal smooth muscle cells in atherosclerosis-prone or atherosclerosis-resistant regions of the human aorta.
        J Vasc Res. 2008; 45: 303-313
        • Hwang J.
        • Saha A.
        • Boo Y.C.
        • et al.
        Oscillatory shear stress stimulates endothelial production of O2 from p47phox-dependent NAD(P)H oxidases, leading to monocyte adhesion.
        J Biol Chem. 2003; 278: 47291-47298
        • Melchionna R.
        • Porcelli D.
        • Mangoni A.
        • et al.
        Laminar shear stress inhibits CXCR4 expression on endothelial cells: functional consequences for atherogenesis.
        FASEB J. 2005; 19: 629-631
        • Azuma K.
        • Watada H.
        • Niihashi M.
        • et al.
        A new En face method is useful to quantitate endothelial damage in vivo.
        Biochem Biophys Res Commun. 2003; 309: 384-390
        • Caplan B.A.
        • Schwartz C.J.
        Increased endothelial cell turnover in area of in vivo Evans blue uptake in the pig aorta.
        Atherosclerosis. 1973; 17: 401-417
        • Lin S.J.
        • Jan K.M.
        • Schuessler G.
        • Weinbaum S.
        • Chien S.
        Enhanced macromolecular permeability of aortic endothelial cells in association with mitosis.
        Atherosclerosis. 1988; 73: 223-232
        • Garcia-Cardeña G.
        • Comander J.
        • Anderson K.R.
        • Blackman B.R.
        • Gimbrone Jr., M.A.
        Biomechanical activation of vascular endothelium as a determinant of its functional phenotype.
        PNAS. 2001; 98: 4478-4485
        • Sokabe T.
        • Yamamoto K.
        • Ohura N.
        • et al.
        Differential regulation of urokinase-type plasminogen activator expression by fluid shear stress in human coronary artery endothelial cells.
        Am J Physiol Heart Circ Physiol. 2004; 287: H2027-H2034
        • Hagiwara H.
        • Mitsumata M.
        • Yamane T.
        • Jin X.
        • Yoshida Y.
        Laminar shear stress-induced GRO mRNA and protein expression in endothelial cells.
        Circulation. 1998; 98: 2584-2590
        • Uei H.
        • Matsuzaki H.
        • Oda H.
        • Nakajima S.
        • Tokuhashi Y.
        • Esumi M.
        Gene expression changes in an early stage of intervertebral disc degeneration induced by passive cigarette smoking.
        Spine. 2006; 31: 510-514
        • Ritt M.G.
        • Mayor J.
        • Wojcieszyn J.
        • Smith III, R.
        • Barton C.L.
        • Modiano J.F.
        Sustained nuclear localization of p21/WAF-1 upon growth arrest induced by contact inhibition.
        Cancer Lett. 2000; 158: 73-84
        • Nishiyama A.
        • Matsui M.
        • Iwata S.
        • et al.
        Identification of thioredoxin-binding protein-2/vitamin D3 up-regulated protein 1 as a negative regulator of thioredoxin function and expression.
        J Biol Chem. 1999; 274: 21645-21650
        • Yamawaki H.
        • Pan S.
        • Lee R.T.
        • Berk B.C.
        Fluid shear stress inhibits vascular inflammation by decreasing thioredoxin-interacting protein in endothelial cells.
        J Clin Invest. 2005; 115: 733-738
        • Wright H.P.
        Mitosis pattern in aortic endothelium.
        Atherosclerosis. 1972; 15: 93-100
        • Yao Y.
        • Rabodzey A.
        • Dewey Jr., C.F.
        Glycocalyx modulates the motility and proliferative response of vascular endothelium to fluid shear stress.
        Am J Physiol Heart Circ Physiol. 2007; 293: H1023-H1030
        • Chappell D.C.
        • Varner S.E.
        • Nerem R.M.
        • Medford R.M.
        • Alexander R.W.
        Oscillatory shear stress stimulates adhesion molecule expression in cultured human endothelium.
        Circ Res. 1998; 82: 532-539
        • Tsao P.S.
        • Buitrago R.
        • Chan J.R.
        • Cooke J.P.
        Fluid flow inhibits endothelial adhesiveness. Nitric oxide and transcriptional regulation of VCAM-1.
        Circulation. 1996; 94: 1682-1689
        • Dai G.
        • Vaughn S.
        • Zhang Y.
        • Wang E.T.
        • Garcia-Cardena G.
        • Gimbrone Jr., M.A.
        Biomechanical forces in atherosclerosis-resistant vascular regions regulate endothelial redox balance via phosphoinositol 3-kinase/Akt-dependent activation of Nrf2.
        Circ Res. 2007; 101: 723-733
        • Cunningham K.S.
        • Gotlieb A.
        The role of shear stress in the pathogenesis of atherosclerosis.
        Lab Invest. 2005; 85: 9-23
        • O’Reilly M.A.
        Redox activation of p21Cip1/WAF1/Sdi1: a multifunctional regulator of cell survival and death. Forum review.
        Antioxid Redox Signal. 2005; 7: 108-118
        • Condorelli G.
        • Aycock J.K.
        • Frati G.
        • Napoli C.
        Mutated p21/WAF/CIP transgene overexpression reduces smooth muscle cell proliferation, macrophage deposition, oxidation-sensitive mechanisms, and restenosis in hypercholesterolemic apolipoprotein E knockout mice.
        FASEB J. 2001; 15: 2162-2170
        • Zernecke A.
        • Shagdarsuren E.
        • Weber C.
        Chemokines in atherosclerosis: an update.
        Arterioscler Thromb Vasc Biol. 2008; 28: 1897-1908
        • Hillyer P.
        • Mordelet E.
        • Flynn G.
        • Male D.
        Chemokines, chemokine receptors and adhesion molecules on different human endothelia: discriminating the tissue-specific functions that affect leukocyte migration.
        Clin Exp Immunol. 2003; 134: 431-441
        • Jiang M.-Z.
        • Tsukahara H.
        • Ohshima Y.
        • et al.
        Effects of antioxidant and nitric oxide on chemokine production in TNF-α-stimulated human dermal microvascular endothelial cells.
        Free Radic Res. 2004; 38: 473-480
        • Nakamura H.
        • Herzenberg L.A.
        • Bai J.
        • et al.
        Circulating thioredoxin suppresses lipopolysaccharide-induced neutrophil chemotaxis.
        PNAS USA. 2001; 98: 15143-15148