NHERF4 hijacks Mas-mediated PLC/AKT signaling to suppress the invasive potential of clear cell renal cell carcinoma cells
NHERF4 hijacks Mas-mediated PLC/AKT signaling to suppress the invasive potential of clear cell renal cell carcinoma cells
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NHERF4 劫持 Mas 介导的 PLC/AKT 信号传导抑制透明细胞肾细胞癌细胞的侵袭潜力
DOI：
10.1016/j.canlet.2021.06.021
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发表时间：
2021-07-05
期刊：
影响因子：
9.7
通讯作者：
He，Junqi
中科院分区：
文献类型：
--
作者：
Yang，Ying;Liang，Jihuan;He，Junqi
据报道，Mas受体通过依赖Ang-(1-7)的AKT信号传导促进透明细胞肾细胞癌（ccRCC）细胞的迁移和侵袭。然而，Mas功能调控的机制尚不清楚。本研究利用表面等离子体共振（SPR）和质谱（MS）技术鉴定了8种含PDZ结构域的蛋白为Mas相互作用体。NHERF4是多个独立ccRCC数据集中唯一下调的基因。GST下拉和共免疫沉淀实验证实了NHERF4和Mas之间的物理相互作用。通过NHERF4过表达和敲低实验，我们发现NHERF4抑制mas诱导的ccRCC细胞的迁移、侵袭和内转移。在机制上，NHERF4抑制mas刺激的AKT磷酸化和PLC/Ca2+反应。我们进一步证明，NHERF4通过调控PLC/AKT信号轴，抑制mas介导的ccRCC细胞迁移和侵袭。对ccRCC数据的分析显示，NHERF4低水平表达与较高的TNM分期相关，并独立预测ccRCC患者预后不良。总的来说，我们的研究确定了NHERF4是ccRCC侵袭性的一种新的调节因子，也是一种预后生物标志物，可能有助于确定ccRCC患者的最佳治疗策略。
The Mas receptor has been reported to promote migration and invasion of clear cell renal cell carcinoma (ccRCC) cells via Ang-(1–7)-dependent AKT signaling. However, the mechanism underlying the regulation of Mas function remains unknown. Here, eight PDZ domain-containing proteins were identified as Mas interactors using surface plasmon resonance (SPR) coupled to mass spectrometry (MS). NHERF4 was the only downregulated gene across multiple independent ccRCC datasets. GST pull-down and co-immunoprecipitation assays confirmed physical interaction between NHERF4 and Mas. Using NHERF4 overexpression and knockdown assays, we found that NHERF4 inhibited Mas-induced migration, invasion andin vivometastasis of ccRCC cells. Mechanistically, NHERF4 suppressed Mas-stimulated AKT phosphorylation and the PLC/Ca2+response. We further demonstrated that NHERF4 compromised Mas-mediated migration and invasion of ccRCC cells via regulation of the PLC/AKT signaling axis. Analysis of the ccRCC dataset revealed that low levels of NHERF4 expression were correlated with higher TNM stage, and independently predicted poor prognosis of ccRCC patients. Overall, our study identified NHERF4 as a novel regulator of ccRCC invasiveness, and a prognostic biomarker, which may be beneficial for determining optimal therapeutic strategies for ccRCC patients.