Blockage of store-operated Ca(2+) entry antagonizes Epstein-Barr virus-promoted angiogenesis by inhibiting Ca(2+) signaling-regulated VEGF production in nasopharyngeal carcinoma.
Blockage of store-operated Ca(2+) entry antagonizes Epstein-Barr virus-promoted angiogenesis by inhibiting Ca(2+) signaling-regulated VEGF production in nasopharyngeal carcinoma.
复制标题
在鼻咽癌中，阻断钙池操纵的 Ca(2) 进入可通过抑制 Ca(2) 信号调节的 VEGF 产生来拮抗 Epstein-Barr 病毒促进的血管生成。
DOI：
10.2147/cmar.s159441
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发表时间：
2018
影响因子：
3.3
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Ye J;Huang J;He Q;Zhao W;Zhou X;Zhang Z;Li Y;Wei J;Zhang J
EB病毒（EBV）通过使鼻咽癌（NPC）细胞获得其恶性生物学行为所需的各种能力，积极促进鼻咽癌（NPC）的病理过程。我们早期的研究表明，EBV 编码的潜膜蛋白 1 (LMP1) 通过在细胞外表皮生长因子 (EGF) 刺激下促进钙库操纵的 Ca2+ 进入 (SOCE) 来增强血管内皮生长因子 (VEGF) 介导的血管生成。然而，必须在体内适当评估 SOCE 阻断对 EBV 促进的血管生成的拮抗作用，并且 EBV 感染对 EGF 引发的胞质 Ca2+ 信号传导的整体影响仍有待进一步阐明，该信号调节 VEGF 介导的血管生成。本研究采用了两种 EBV 感染的 NPC 细胞系 CNE2-EBV 和 HK1-EBV 及其亲代细胞系。在单个荧光 Ca2+ 指示剂负载细胞中测量动态胞质 Ca2+ 变化。通过酶联免疫吸附测定（ELISA）测定VEGF的产生量。作为体外血管生成测定，对人脐静脉内皮细胞（HUVEC）形成的管网络进行定量评估。利用同时携带 EBV 阳性/阴性异种移植物的小鼠模型来评估体内肿瘤生长和血管生成。 EBV 感染可靠地促进移植肿瘤生长，同时增强血管生成。将EBV引入EBV阴性NPC细胞增加了EGF刺激的VEGF产生，同时放大了EGF诱发的Ca2+反应。使用特异性 SOCE 抑制剂 2-氨基乙基二苯硼酸酯 (2-APB) 抑制 EBV 促进的 Ca2+ 信号传导，可在体外有效拮抗 EBV 促进的 VEGF 产生和内皮管形成。 SOCE 的药理学阻断在 EBV 阳性异种移植物中表现出抗血管生成作用。 SOCE 可以作为治疗 NPC 的候选药理学靶点，因为通过 SOCE 阻断 Ca2+ 信号传导是抑制 EBV 驱动的 NPC 细胞恶性特征的可行策略。
Epstein–Barr virus (EBV) actively contributes to the pathological process of nasopharyngeal carcinoma (NPC) by enabling NPC cells to acquire various capacities required for their malignant biological actions. Our earlier works demonstrated that EBV-encoded latent membrane protein 1 (LMP1) enhanced vascular endothelial growth factor (VEGF)-mediated angiogenesis by boosting store-operated Ca2+ entry (SOCE) upon extracellular epidermal growth factor (EGF) stimulation. However, the antagonistic effects of SOCE blockage on EBV-promoted angiogenesis must be appropriately evaluated in vivo, and the global effect of EBV infection on the EGF-elicited cytosolic Ca2+ signaling, which regulates VEGF-mediated angiogenesis remains to be further clarified. Two EBV-infected NPC cell lines, CNE2-EBV and HK1-EBV, along with their parental cell lines were employed in the present study. Dynamic cytosolic Ca2+ changes were measured in individual fluorescent Ca2+ indicator-loaded cells. Amounts of VEGF production were determined by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs)-formed tube networks were quantitatively evaluated as an in vitro angiogenesis assay. A mouse model concurrently bearing EBV-positive/negative xenografts was utilized to evaluate the tumor growth and angiogenesis in vivo. EBV infection reliably promoted transplanted tumor growth while enhancing angiogenesis. Introduction of EBV into EBV-negative NPC cells increased the EGF-stimulated VEGF production while amplifying the EGF-evoked Ca2+ responses. Inhibition of the EBV-boosted Ca2+ signaling using 2-aminoethyl diphenylborinate (2-APB), a specific SOCE inhibitor, effectively antagonized the EBV-promoted VEGF production and endothelial tube formation in vitro. Pharmacological blockage of SOCE exhibited anti-angiogenic effect in the EBV-positive xenografts. SOCE can serve as a candidate pharmacological target for treating NPC, as blockage of the Ca2+ signaling via SOCE is a feasible strategy to suppress the EBV-driven malignant profiles in NPC cells.