Recruitment of Bone Marrow-Derived Endothelial Progenitor Cells to Vascular Endothelium Involves E-Selectin Dependent Mechanism

Nishiwaki, Yasunobu; Yoshida, Masayuki; Iwaguro, Hideki; Masuda, Haruchika; Asahara, Takayuki; Isobe, Mitsuaki

doi:10.1016/j.carpath.2004.03.515

Cited by 1 publication

(4 citation statements)

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“…Current results compliment this work and emphasise the efficacy of siRNA in the field of vascular biology. We have also reinforced the work of Nishiwaki et al (using antibodies to downregulate E-selectin) and Vajkoczy et al (using an in vivo method to silence Eselectin expression) in demonstrating that E-selectin is important in EPC adhesion by using a different method (siRNA transfection) [6,7].…”

Section: Discussionsupporting

confidence: 77%

“…For use in the flow-chamber apparatus, HUVECs were plated onto 22 mm fibronectin-coated glass coverslips as has been described previously [13,14]. Preparation of endothelial progenitor cells was previously described in detail [7].…”

Section: Cell Culture and Reagentsmentioning

confidence: 99%

“…Western blot analysis was performed using lysates prepared from HUVEC, as described previously [7]. An equal amount of protein (10 μg) from each condition was subjected to 5-20% SDS-PAGE.…”

Section: Western Blotmentioning

confidence: 99%

“…We recently reported an important role of endothelial E-selectin in mediating endothelial progenitor cell (EPC) recruitment [7].…”

Section: Introductionmentioning

confidence: 99%

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Gene silencing of E-selectin block recruitment of endothelial progenitor cell to vascular endothelium under flow

Sharma¹,

Yoshida²

2010

JBiSE

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Short interfering RNA (siRNA) is a powerful technique that can suppress gene expression in a variety of cells including mammalian cells. Endothelial progenitor cells (EPCs) are bone marrow—derived haematopoietic progenitor cells that have been implicated in vasculogenesis. We demonstrated for the first time that gene silencing of endothelial E-selectin using siRNA transfection in human umbilical vein endothelial cells (HUVECs) causes inhibition of EPC adhesion under flow conditions. Fluorescence immunobinding assay analysis showed that significant reduction of E-selectin surface expression in HUVECs (activated with IL-1β (10 U/mL) for 4 h) transfected with siRNA against E-selectin, but not in HUVECs transfected with LacZ siRNA (control). An EPC adhesion assay under flow conditions (shear stress = 1.0 dyne/cm2) then demonstrated that HUVECs transfected with E-selectin siRNA supported significantly less adhesion of EPCs than those HUVECs treated with control siRNA and no siRNA after activation by IL-1β (p < 0.05). Our experiments have shown the importance of E-selectin in EPC adhesion to HUVECs and the potential utility of gene silencing of E-selectin in EPC recruitment

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Section: Discussionsupporting

confidence: 77%