NEK9, a novel effector of IL-6/STAT3, regulates metastasis of gastric cancer by targeting ARHGEF2 phosphorylation.
NEK9, a novel effector of IL-6/STAT3, regulates metastasis of gastric cancer by targeting ARHGEF2 phosphorylation.
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NEK9是IL-6/STAT3的新型效应子，通过靶向ARHGEF2磷酸化调节胃癌转移
DOI：
10.7150/thno.53169
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发表时间：
2021
期刊：
影响因子：
12.4
通讯作者：
Shang Y
中科院分区：
文献类型：
--
作者：
Lu G;Tian S;Sun Y;Dong J;Wang N;Zeng J;Nie Y;Wu K;Han Y;Feng B;Shang Y
基本原理：来自肿瘤微环境的炎症刺激在癌症进展中起重要作用。然而，炎症促进胃癌转移的机制尚不清楚。方法：通过体外功能丧失和功能获得实验以及转移动物模型验证NEK 9的作用。GST pull-down检测细胞骨架重组相关分子。通过磷酸化蛋白质组学分析、免疫沉淀（IP）和体外激酶试验研究NEK 9对ARHGEF 2的调节作用。利用荧光素酶报告基因和染色质免疫沉淀技术（ChIP）研究了miR-520 f-3 p的转录调控。免疫组化法检测胃癌组织中这些蛋白的表达。结果：NEK 9直接调节胃癌细胞的运动和RhoA的激活。NEK 9对ARHGEF 2的磷酸化是这一过程的关键步骤。NEK 9是miR-520 f-3 p的直接靶点，miR-520 f-3 p通过IL-6介导的STAT 3活化而被转录抑制。miR-520 f-3 p的减少通过靶向GP 130导致IL-6/STAT 3的扩增。在淋巴结和远处转移灶中证实NEK 9、GP 130和p-STAT 3水平同时升高。NEK 9、GP 130和STAT 3的增加与GC患者的总生存率降低相关。结论：这项研究表明，IL-6激活STAT 3可在转录上抑制miR-520 f-3 p，并减弱miR-520 f-3 p对NEK 9和GP 130的抑制作用。GP 130的增加增强了这种信号传导，NEK 9通过靶向ARHGEF 2的磷酸化直接影响细胞运动和RhoA活化。靶向IL-6-STAT 3-NEK 9通路可能是GC治疗的新策略。
Rationale: Inflammatory stimuli from the tumor microenvironment play important roles in cancer progression. However, the mechanism of promotion of cancer metastasis by inflammation in gastric cancer (GC) is poorly understood. Methods: The roles of NEK9 were validated via loss-of-function and gain-of-function experiments in vitro and in an animal model of metastasis. Cytoskeletal reorganization-associated molecules were detected by GST pull-down. The regulation of ARHGEF2 by NEK9 was investigated by phosphoproteomics analysis, immunoprecipitation (IP) and in vitro kinase assay. The transcriptional regulation of miR-520f-3p was studied using luciferase reporter and chromatin immunoprecipitation (ChIP). The expression of these proteins in GC tissues was examined by immunohistochemistry. Results: NEK9 directly regulates cell motility and RhoA activation in GC. The phosphorylation of ARHGEF2 by NEK9 is the key step of this process. NEK9 is a direct target of miR-520f-3p, which is transcriptionally suppressed by IL-6-mediated activation of STAT3. A decrease in miR-520f-3p leads to the amplification of IL-6/STAT3 by targeting GP130. A simultaneous elevation of the levels of NEK9, GP130 and p-STAT3 was confirmed in the lymph nodes and distant metastases. An increase in NEK9, GP130 and STAT3 is associated with reduced overall survival of GC patients. Conclusion: This study demonstrates that activation of STAT3 by IL-6 transcriptionally suppresses miR-520f-3p and diminishes the inhibitory effects of miR-520f-3p on NEK9 and GP130. An increase in GP130 enhances this signaling, and NEK9 directly influences cell motility and RhoA activation by targeting the phosphorylation of ARHGEF2. Targeting the IL-6-STAT3-NEK9 pathway may be a new strategy for GC treatment.