Differential expression of functional nucleoside transporters in non-differentiated and differentiated human endothelial progenitor cells

Guzman-Gutierrez, E; Sandoval C.; NOVA, E; CASTILLO, JL; Vera, JC; Lamperti, L; Krause, B; Salomon, C; Sepulveda, C; Aguayo, C.; Sobrevia L.

Abstract

Extracellular adenosine removal is via human equilibrative nucleoside transporters 1 (hENT1) and 2 (hENT2) in the endothelium, thus regulating adenosine-induced revascularization and angiogenesis. Since human endothelial progenitor cells (hEPCs) promote revascularization, we hypothesize differential expression of nucleoside transporters in hEPCs. hEPCs were cultured 3 (hEPC-3d) or 14 (hEPC-14d) days. RT-PCR for prominin 1, CD34, octamer-4, kinase insert domain receptor, oxidized low-density lipoprotein (lectin-like) receptor 1 and tyrosine endothelial kinase was used to evaluate phenotypic differentiation. Flow cytometry was used to estimate CD34+/KDR- (non-differentiated), CD34-/KDR+ (differentiated) or CD34+/KDR+ (mixed) cell populations. Adenosine transport was measured in absence or presence of sodium, S-(4-nitrobenzyl)-6-thio-inosine (NBTI, 1-10 μM), inosine, hypoxanthine or guanine (0.1-5 mM), hENTs protein abundance by western blot, and hENTs, hCNT1, hCNT2 and hCNT3 mRNA expression by real time RT-PCR. hEPC-3d cells were CD34+/KDR- compared with hEPC-14d cells that were CD34-/KDR+. hEPC-3d cells exhibit hENT1-like adenosine transport (NBTI-sensitive, Na+- independent), which is absent in hEPC-14d cells. hEPC-14d cells exhibit two transport components: component 1 (NBTI insensitive, Na+-independent) and component 2 (NBTI insensitive, Na+-dependent, Hill coefficient ∼1.8), the latter resembling CNT3-like transport. hEPC-3d cells express hENT1 protein and mRNA, which is reduced (∼90%) in hEPC-14d cells, but instead only hCNT3 mRNA is expressed in this cell type. hENT2, hCNT1 and hCNT2 were undetectable in hEPCs. Thus, hEPCs exhibit a differential expression of hENT1 and hCNT3 functional nucleoside transporters, which could be related with its differentiation stage. © 2010 Elsevier Ltd. All rights reserved.

Más información

Título según WOS: Differential expression of functional nucleoside transporters in non-differentiated and differentiated human endothelial progenitor cells
Título según SCOPUS: Differential expression of functional nucleoside transporters in non-differentiated and differentiated human endothelial progenitor cells
Título de la Revista: PLACENTA
Volumen: 31
Número: 10
Editorial: W. B. Saunders Co., Ltd.
Fecha de publicación: 2010
Página de inicio: 928
Página final: 936
Idioma: English
URL: http://linkinghub.elsevier.com/retrieve/pii/S0143400410002857
DOI:

10.1016/j.placenta.2010.07.016

Notas: ISI, SCOPUS