Regulation of store-operated Ca2+ entry activity by cell cycle dependent up-regulation of Orai2 in brain capillary endothelial cells.
Regulation of store-operated Ca2+ entry activity by cell cycle dependent up-regulation of Orai2 in brain capillary endothelial cells.
复制标题
通过脑毛细血管内皮细胞中细胞周期依赖性上调 Orai2 来调节钙池操纵的 Ca2 进入活性。
DOI：
10.1016/j.bbrc.2015.02.127
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发表时间：
2015
影响因子：
3.1
通讯作者：
Yuji Imaizumi
中科院分区：
文献类型：
--
作者：
Hiroaki Kito;Hisao Yamamura;Yoshiaki Suzuki;Susumu Ohya;Kiyofumi Asai;Yuji Imaizumi
通过Orai1和STIM1复合物的钙库操纵Ca2+进入（SOCE）被认为在血管内皮细胞的细胞功能调节中具有必然的作用，而关于Orai2的贡献却知之甚少。定量 PCR 和蛋白质印迹分析表明，在牛脑毛细血管内皮细胞系 t-BBEC117 中，除了 Orai1 和 STIM1 之外，还表达了 Orai2 和 STIM2。在 t-BBEC117 的指数生长过程中，Orai1 和 STIM1 的敲低显着降低了 SOCE 活性，而 Orai2 和 STIM2 siRNA 则没有影响。为了检查内源性 SOCE 活性是否有助于细胞周期进程的调节，使用双胸苷阻断来同步 t-BBEC117。在 G2/M 期，与 G0/G1 期相比，通过 SOCE 的 Ca2+ 内流减少，Orai2 表达增加。当Orai2在G2/M期被敲低时，SOCE的减少被消除，细胞增殖部分减弱。总的来说，Orai1 通过功能表达显着促进细胞增殖，这可能与细胞周期阶段无关。在构建中，Orai2 在 G2/M 期特异性上调，负向调节 SOCE 活性，并可能有助于调节脑毛细血管内皮细胞的细胞周期进程。
Store-operated Ca2+entry (SOCE) via Orai1 and STIM1 complex is supposed to have obligatory roles in the regulation of cellular functions of vascular endothelial cells, while little is known about the contribution of Orai2. Quantitative PCR and Western blot analyses indicated the expression of Orai2 and STIM2, in addition to Orai1 and STIM1 in bovine brain capillary endothelial cell line, t-BBEC117. During the exponential growth of t-BBEC117, the knockdown of Orai1 and STIM1 significantly reduced the SOCE activity, whereas Orai2 and STIM2 siRNAs had no effect. To examine whether endogenous SOCE activity contributes to the regulation of cell cycle progression, t-BBEC117 were synchronized using double thymidine blockage. At the G2/M phase, Ca2+influx via SOCE was decreased and Orai2 expression was increased compared to the G0/G1 phase. When Orai2 was knocked down at the G2/M phase, the decrease in SOCE was removed, and cell proliferation was partly attenuated. Taken together, Orai1 significantly contributes to cell proliferation via the functional expression, which is presumably independent of the cell cycle phases. In construct, Orai2 is specifically up-regulated during the G2/M phase, negatively modulates the SOCE activity, and may contribute to the regulation of cell cycle progression in brain capillary endothelial cells.