Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation.
Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation.
复制标题
DOI：
10.1161/circresaha.111.243352
复制
发表时间：
2011-05-13
影响因子：
20.1
通讯作者：
Beech DJ
中科院分区：
文献类型：
--
作者：
Li J;Cubbon RM;Wilson LA;Amer MS;McKeown L;Hou B;Majeed Y;Tumova S;Seymour VA;Taylor H;Stacey M;O'Regan D;Foster R;Porter KE;Kearney MT;Beech DJ
在免疫系统中发现了 Orai1 和相关的钙释放激活钙 (CRAC) 通道。已提出在内皮细胞中也存在，但与内皮生物学的相关性大多未知。本研究的目的是探讨Orai1和CRAC通道与血管内皮生长因子（VEGF）和内皮管形成的相关性。在人脐静脉内皮细胞中，短干扰RNA或显性失活突变体Orai1破坏Orai1可抑制钙释放激活（钙池操作）的钙内流、VEGF诱发的钙内流、细胞迁移和体外管形成。外源野生型Orai1 的表达挽救了管的形成。 VEGF 受体 2 和 Orai1 部分共定位。 Orai1 破坏还抑制人血液内皮祖细胞的钙进入和管形成。一种已知的免疫细胞 CRAC 通道阻滞剂（3-氟吡啶-4-羧酸（2',5'-二甲氧基联苯-4-基）酰胺）是内皮细胞中钙库操纵的钙进入的强阻滞剂，并抑制 VEGF 在三种类型的人内皮细胞中引起的钙进入。该化合物对 VEGF 诱发的钙释放、STIM1 簇以及两种类型的内皮 TRP 通道（TRPC6 和 TRPV4）缺乏影响。该化合物在体外抑制人内皮细胞迁移和管形成，并在体内抑制鸡绒毛尿囊膜中的血管生成，而不影响细胞活力。与免疫细胞钙进入相比，该化合物对内皮细胞的效力高出 100 倍。数据表明 Orai1 和 CRAC 通道在 VEGF 诱发的钙进入中发挥积极作用，并为化学调节血管生成提供了新的机会。
Orai1 and the associated calcium release-activated calcium (CRAC) channel were discovered in the immune system. Existence also in endothelial cells has been suggested but the relevance to endothelial biology is mostly unknown. The aim of this study was to investigate the relevance of Orai1 and CRAC channels to vascular endothelial growth factor (VEGF) and endothelial tube formation. In human umbilical vein endothelial cells Orai1 disruption by short-interfering RNA or dominant-negative mutant Orai1 inhibited calcium-release-activated (store-operated) calcium entry, VEGF-evoked calcium-entry, cell migration, and in vitro tube formation. Expression of exogenous wild-type Orai1 rescued the tube formation. VEGF receptor-2 and Orai1 partially co-localized. Orai1 disruption also inhibited calcium-entry and tube formation in endothelial progenitor cells from human blood. A known blocker of the immune cell CRAC channel (3-fluoropyridine-4-carboxylic acid (2′,5′-dimethoxybiphenyl-4-yl)amide) was a strong blocker of store-operated calcium entry in endothelial cells and inhibited calcium-entry evoked by VEGF in three types of human endothelial cell. The compound lacked effect on VEGF-evoked calcium-release, STIM1 clustering, and two types of endothelial TRP channel, TRPC6 and TRPV4. Without effect on cell viability the compound inhibited human endothelial cell migration and tube formation in vitro and suppressed angiogenesis in vivo in the chick chorioallantoic membrane. The compound showed 100-fold greater potency for endothelial compared with immune cell calcium entry. The data suggest positive roles for Orai1 and CRAC channels in VEGF-evoked calcium entry and new opportunity for chemical modulation of angiogenesis.