β-Arrestin 1 is required for endothelin-1-induced NF-κB activation in ovarian cancer cells
β-Arrestin 1 is required for endothelin-1-induced NF-κB activation in ovarian cancer cells
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DOI：
10.1016/j.lfs.2014.01.078
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发表时间：
2014-11-24
期刊：
影响因子：
6.1
通讯作者：
Rosano, Laura
中科院分区：
文献类型：
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作者：
Cianfrocca, Roberta;Tocci, Piera;Rosano, Laura
目的：在上皮性卵巢癌（EOC）中，表皮覆盖素-1 (ET-1)/内皮素A受体（ETAR）信号的激活与许多肿瘤促进作用有关，如增殖、血管生成、侵袭和转移。这些作用依赖于关键信号通路的激活，如MAPK、Akt和β -连环蛋白，通过β -阻滞蛋白1 （β -arr1）的特异性细胞质和核支架功能。在这里，我们评估了ET-1/ETAR通过β -ar -1募集促进EOC细胞NF-kappa B信号传导的潜在作用。主要方法：我们使用在ET-1和ETAR拮抗剂BQ123存在或不存在的情况下培养的HEY EOC细胞。免疫印迹法检测p65和I κ pa- b α的磷酸化水平。通过核提取物免疫沉淀实验评价p65与β -arr1的相互作用。通过转染NF-kappa B驱动的荧光素酶报告结构来评估NF-kappa B启动子活性。通过shRNA沉默β -arr1和表达β -arr1- flag表达载体来评估β -arr1的功能。关键发现：在EOC细胞中，ET-1促进p65亚基和细胞质抑制剂I kappa B的磷酸化，进而导致NF-kappa B转录活性增加。这些作用被BQ123和β - ar1沉默所抑制，这表明ET-1通过ETAR促进β -arr1的募集来调节NF-kappa B信号传导。此外，p65和β -arr1之间的核物理相互作用表明β -arr1在etar驱动的NF-kappa B转录活性中具有核功能。意义：总的来说，这些发现揭示了一个以前未被认识的途径，该途径依赖于β -arr1来维持NF-kappa B信号传导，以响应卵巢癌中ETAR的激活。(C) 2014年作者。Elsevier Inc.出版。这是一篇基于CC BY-NC-ND许可（http://creativecommons.org/licenses/by-nc-nd/3.0/）的开放获取文章。
Aims: In epithelial ovarian cancer (EOC), activation of enclothelin-1 (ET-1)/endothelin A receptor (ETAR) signalling is linked to many tumor promoting effects, such as proliferation, angiogenesis, invasion and metastasis. These effects are dependent by the activation of critical signalling pathways, such as MAPK, Akt, and beta-catenin, through specific cytosolic and nuclear scaffolding functions of beta-arrestin 1 (beta-arr1). Here, we have assessed the potential role of ET-1/ETAR in promoting NF-kappa B signalling in EOC cells through beta-arr-1 recruitment.Main methods: We used cultured HEY EOC cells cultured in the presence or absence of ET-1 and the ETAR antagonist BQ123. The phosphorylation of p65 and I kappa-B alpha was evaluated by immunoblotting analysis. The interaction between p65 and beta-arr1 was evaluated by immunoprecipitation experiments in nuclear extracts. NF-kappa B promoter activity was evaluated by transfection with NF-kappa B-driven luciferase reporter construct. Assessment of the function of beta-arr1 was achieved by beta-arr1 silencing with shRNA and expression of beta-arr1-FLAG expression vector.Key findings: In EOC cells, ET-1 promotes the phosphorylation of p65 subunit and the cytoplasmic inhibitor I kappa B that in turn led to increased NF-kappa B transcriptional activity. These effects were inhibited by the use of BQ123, as well as by beta-arr-1 silencing, suggesting that ET-1 through ETAR promotes the recruitment of beta-arr1 to regulate NF-kappa B signalling. Moreover, the nuclear physical interaction between p65 and beta-arr1 indicates a nuclear function of beta-arr-1 in ETAR-driven NF-kappa B transcriptional activity.Significance: Altogether these findings reveal a previously unrecognized pathway that depends on beta-arr1 to sustain NF-kappa B signalling in response to ETAR activation in ovarian cancer. (C) 2014 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).