Store-operated Ca2+ entry is remodelled and controls in vitro angiogenesis in endothelial progenitor cells isolated from tumoral patients.
Store-operated Ca2+ entry is remodelled and controls in vitro angiogenesis in endothelial progenitor cells isolated from tumoral patients.
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DOI：
10.1371/journal.pone.0042541
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Porta C
中科院分区：
文献类型：
--
作者：
Lodola F;Laforenza U;Bonetti E;Lim D;Dragoni S;Bottino C;Ong HL;Guerra G;Ganini C;Massa M;Manzoni M;Ambudkar IS;Genazzani AA;Rosti V;Pedrazzoli P;Tanzi F;Moccia F;Porta C
内皮祖细胞(EPC)可以从骨髓中招募，以维持肿瘤的血管形成并促进转移开关。了解驱动内皮祖细胞增殖和小管生成的分子机制可以为替代抗血管生成治疗勾勒出新的靶点。内质网钙离子通道STIM1和质膜钙通道Orai1之间的相互作用是调节人内皮祖细胞生长的重要机制。由于肿瘤的发生可能与肿瘤细胞在钙离子内流时独立生长的能力有关，因此评估SOCE是否也调节肿瘤患者EPC依赖的血管生成是很重要的。本研究采用钙离子成像、膜下和线粒体重组Aequorin、实时聚合酶链式反应、基因沉默技术和蛋白质印迹分析等方法，研究了肾癌患者外周血中SOCE的表达和作用，并与对照组(N-EPC)进行了比较。在RCC-EPC中，SOCE被药物刺激(即环匹阿松酸)或生理刺激(即ATP)激活，并且对BTP-2和三价阳离子La3+和Gd3+选择性敏感。此外，2-APB在低浓度时增强了thapsigarin引起的SOCE，而较高剂量则抑制了SOCE。相反，抗血管生成药物羧胺三氮唑(CaI)可阻断SOCE和细胞内钙离子的释放。SOCE与Orai1、STIM1和瞬时受体电位通道1(TRPC1)在mRNA和蛋白水平的过度表达有关。细胞内钙缓冲、BAPTA、BTP-2和CA1抑制RCC-EPC的增殖和小管生成。STIM1、Orai1和TRPC1的基因抑制阻断了CPA诱导的肾癌内皮祖细胞的SOCE。SOCE在肾癌患者的EPC中重塑，并因其控制增殖和小管生成的能力而成为干扰RCC血管形成的新的分子靶点。
Endothelial progenitor cells (EPCs) may be recruited from bone marrow to sustain tumor vascularisation and promote the metastatic switch. Understanding the molecular mechanisms driving EPC proliferation and tubulogenesis could outline novel targets for alternative anti-angiogenic treatments. Store-operated Ca2+ entry (SOCE), which is activated by a depletion of the intracellular Ca2+ pool, regulates the growth of human EPCs, where is mediated by the interaction between the endoplasmic reticulum Ca2+-sensor, Stim1, and the plasmalemmal Ca2+ channel, Orai1. As oncogenesis may be associated to the capability of tumor cells to grow independently on Ca2+ influx, it is important to assess whether SOCE regulates EPC-dependent angiogenesis also in tumor patients. The present study employed Ca2+ imaging, recombinant sub-membranal and mitochondrial aequorin, real-time polymerase chain reaction, gene silencing techniques and western blot analysis to investigate the expression and the role of SOCE in EPCs isolated from peripheral blood of patients affected by renal cellular carcinoma (RCC; RCC-EPCs) as compared to control EPCs (N-EPCs). SOCE, activated by either pharmacological (i.e. cyclopiazonic acid) or physiological (i.e. ATP) stimulation, was significantly higher in RCC-EPCs and was selectively sensitive to BTP-2, and to the trivalent cations, La3+ and Gd3+. Furthermore, 2-APB enhanced thapsigargin-evoked SOCE at low concentrations, whereas higher doses caused SOCE inhibition. Conversely, the anti-angiogenic drug, carboxyamidotriazole (CAI), blocked both SOCE and the intracellular Ca2+ release. SOCE was associated to the over-expression of Orai1, Stim1, and transient receptor potential channel 1 (TRPC1) at both mRNA and protein level The intracellular Ca2+ buffer, BAPTA, BTP-2, and CAI inhibited RCC-EPC proliferation and tubulogenesis. The genetic suppression of Stim1, Orai1, and TRPC1 blocked CPA-evoked SOCE in RCC-EPCs. SOCE is remodelled in EPCs from RCC patients and stands out as a novel molecular target to interfere with RCC vascularisation due to its ability to control proliferation and tubulogenesis.
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影响因子：
5.6
作者：
Bomben, Valerie C.;Turner, Kathryn L.;Barclay, Tia-Tabitha C.;Sontheimer, Harald
通讯作者：
Sontheimer, Harald
影响因子：
5.6
作者：
Ciarcia, Roberto;d'Angelo, Danila;Giordano, Antonio
通讯作者：
Giordano, Antonio
影响因子：
4
作者：
Feldman, Ben;Fedida-Metula, Shlomit;Fishman, Daniel
通讯作者：
Fishman, Daniel
影响因子：
4.8
作者：
Antigny, Fabrice;Girardin, Nathalie;Frieden, Maud
通讯作者：
Frieden, Maud
影响因子：
4
作者：
Harper, Matthew T.;Poole, Alastair W.
通讯作者：
Poole, Alastair W.