GSK-3β Governs Inflammation-Induced NFATc2 Signaling Hubs to Promote Pancreatic Cancer Progression.
GSK-3β Governs Inflammation-Induced NFATc2 Signaling Hubs to Promote Pancreatic Cancer Progression.
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DOI：
10.1158/1535-7163.mct-15-0309
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发表时间：
2016-03
影响因子：
5.7
通讯作者：
Neesse A
中科院分区：
文献类型：
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作者：
Baumgart S;Chen NM;Zhang JS;Billadeau DD;Gaisina IN;Kozikowski AP;Singh SK;Fink D;Ströbel P;Klindt C;Zhang L;Bamlet WR;Koenig A;Hessmann E;Gress TM;Ellenrieder V;Neesse A
我们的目的是研究GSK-3β在胰腺癌中对促炎致癌转录因子NFATc 2的调节和激活的机制、功能和治疗作用。免疫组织化学，定量实时聚合酶链反应，免疫印迹，免疫荧光显微镜和增殖试验被用来分析小鼠和人类组织和细胞系。蛋白质-蛋白质相互作用和启动子调控进行了分析，免疫共沉淀，DNA下拉，报告，和ChIP测定。使用多种胰腺癌细胞系、异种移植物和基因工程小鼠模型（GEMM）进行临床前测定。GSK-3β依赖性SP2磷酸化介导胰腺癌细胞核中NFATc 2蛋白稳定性，刺激胰腺癌生长除了蛋白质稳定外，GSK-3β还通过一种独特的机制维持NFATc 2激活，该机制涉及独立于SP2磷酸化的NFATc 2-STAT 3复合物的稳定。对于NFATc 2-STAT 3复合物的形成，需要GSK-3β介导的STAT 3在Y 705处的磷酸化来刺激NFAT靶启动子（如细胞周期蛋白依赖性激酶-6（CDK-6））的常染色质形成，从而促进肿瘤生长。最后，临床前实验表明，靶向NFATc 2-STAT 3-GSK-3β模块可抑制KrasG 12 D小鼠的增殖、肿瘤生长，并干扰炎症诱导的胰腺癌进展。总之，我们描述了GSK-3β微调NFATc 2和STAT 3转录网络以整合上游信号事件的新机制，这些事件控制胰腺癌的进展和生长。此外，GSK-3β的治疗潜力首次在胰腺癌的相关Kras和炎症诱导的GEMM中得到证实。
We aimed to investigate the mechanistic, functional and therapeutic role of GSK-3β in the regulation and activation of the pro-inflammatory oncogenic transcription factor NFATc2 in pancreatic cancer. Immunohistochemistry, quantitative real-time polymerase chain reaction, immunoblotting, immunofluorescence microscopy and proliferation assays were used to analyze mouse and human tissues and cell lines. Protein-protein interactions and promoter regulation were analyzed by co-immunoprecipitation, DNA pull-down, reporter, and ChIP assays. Preclinical assays were performed using a variety of pancreatic cancer cells lines, xenografts and a genetically engineered mouse model (GEMM). GSK-3β-dependent SP2 phosphorylation mediates NFATc2 protein stability in the nucleus of pancreatic cancer cells stimulating pancreatic cancer growth. In addition to protein stabilization, GSK-3β also maintains NFATc2 activation through a distinct mechanism involving stabilization of NFATc2-STAT3 complexes independent of SP2 phosphorylation. For NFATc2-STAT3 complex formation, GSK-3β-mediated phosphorylation of STAT3 at Y705 is required to stimulate euchromatin formation of NFAT target promoters such as cyclin dependent kinase −6 (CDK-6), which promotes tumor growth. Finally, preclinical experiments suggest that targeting the NFATc2-STAT3-GSK-3β module inhibits proliferation, tumor growth, and interferes with inflammation-induced pancreatic cancer progression in KrasG12D mice. In conclusion, we describe a novel mechanism by which GSK-3β fine-tunes NFATc2 and STAT3 transcriptional networks to integrate upstream signaling events that govern pancreatic cancer progression and growth. Furthermore, the therapeutic potential of GSK-3β is demonstrated for the first time in a relevant Kras and inflammation-induced GEMM for pancreatic cancer.