The microRNA-200/Zeb1 axis regulates ECM-dependent β1-integrin/FAK signaling, cancer cell invasion and metastasis through CRKL.
The microRNA-200/Zeb1 axis regulates ECM-dependent β1-integrin/FAK signaling, cancer cell invasion and metastasis through CRKL.
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DOI：
10.1038/srep18652
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发表时间：
2016-01-05
影响因子：
4.6
通讯作者：
Gibbons DL
中科院分区：
文献类型：
--
作者：
Ungewiss C;Rizvi ZH;Roybal JD;Peng DH;Gold KA;Shin DH;Creighton CJ;Gibbons DL
肿瘤细胞转移是一个复杂的过程，其机制与上皮-间质转化（EMT）有关。microRNA-200家族与Zeb1转录抑制因子之间的双负反馈回路是EMT的主调控因子，但对miR-200如何抑制侵袭的理解尚不完整。我们最近的工作集中在肿瘤细胞活化所必需的肿瘤细胞-基质相互作用上。在这里，我们利用Kras/p53突变小鼠模型和人肺癌细胞系来证明，在miR-200缺失时，整合素β1-胶原I相互作用驱动3D体外迁移/侵袭和体内转移。zeb1依赖性EMT增强肿瘤细胞对ECM成分的反应性，并通过去抑制miR-200的直接靶点CRKL激活FAK/Src通路信号。我们证明了CRKL作为一个适配器分子促进局灶黏附的形成，通过Itgβ1介导由外向内信号驱动细胞侵袭，以及由内向外信号维持细胞侵袭所需的肿瘤细胞-基质接触。重要的是，在泛癌症TCGA分析中，CRKL水平可以预测生存，CRKL敲低可以抑制体内的实验转移，而不影响原发肿瘤的生长。我们的研究结果强调了关键的ecm -肿瘤细胞相互作用由miR-200/ zeb1依赖的EMT调节，激活负责肿瘤细胞侵袭和转移的细胞内信号通路。
Tumor cell metastasis is a complex process that has been mechanistically linked to the epithelial-mesenchymal transition (EMT). The double-negative feedback loop between the microRNA-200 family and the Zeb1 transcriptional repressor is a master EMT regulator, but there is incomplete understanding of how miR-200 suppresses invasion. Our recent efforts have focused on the tumor cell-matrix interactions essential to tumor cell activation. Herein we utilized both our Kras/p53 mutant mouse model and human lung cancer cell lines to demonstrate that upon miR-200 loss integrin β1-collagen I interactions drive 3D in vitro migration/invasion and in vivo metastases. Zeb1-dependent EMT enhances tumor cell responsiveness to the ECM composition and activates FAK/Src pathway signaling by de-repression of the direct miR-200 target, CRKL. We demonstrate that CRKL serves as an adaptor molecule to facilitate focal adhesion formation, mediates outside-in signaling through Itgβ1 to drive cell invasion, and inside-out signaling that maintains tumor cell-matrix contacts required for cell invasion. Importantly, CRKL levels in pan-cancer TCGA analyses were predictive of survival and CRKL knockdown suppressed experimental metastases in vivo without affecting primary tumor growth. Our findings highlight the critical ECM-tumor cell interactions regulated by miR-200/Zeb1-dependent EMT that activate intracellular signaling pathways responsible for tumor cell invasion and metastasis.