α-烯醇化酶（ENO1）控制αv/β3整合素的表达，并调节胰腺癌的黏附、侵袭和转移。
Alpha-enolase (ENO1) controls alpha v/beta 3 integrin expression and regulates pancreatic cancer adhesion, invasion, and metastasis.
作者信息
Principe Moitza, Borgoni Simone, Cascione Mariafrancesca, Chattaragada Michelle Samuel, Ferri-Borgogno Sammy, Capello Michela, Bulfamante Sara, Chapelle Jennifer, Di Modugno Francesca, Defilippi Paola, Nisticò Paola, Cappello Paola, Riganti Chiara, Leporatti Stefano, Novelli Francesco
机构信息
Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.
Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
出版信息
J Hematol Oncol. 2017 Jan 13;10(1):16. doi: 10.1186/s13045-016-0385-8.
BACKGROUND
We have previously shown that in pancreatic ductal adenocarcinoma (PDA) cells, the glycolytic enzyme alpha-enolase (ENO1) also acts as a plasminogen receptor and promotes invasion and metastasis formation. Moreover, ENO1 silencing in PDA cells induces oxidative stress, senescence and profoundly modifies PDA cell metabolism. Although anti-ENO1 antibody inhibits PDA cell migration and invasion, little is known about the role of ENO1 in regulating cell-cell and cell-matrix contacts. We therefore investigated the effect of ENO1 silencing on the modulation of cell morphology, adhesion to matrix substrates, cell invasiveness, and metastatic ability.
METHODS
The membrane and cytoskeleton modifications that occurred in ENO1-silenced (shENO1) PDA cells were investigated by a combination of confocal microscopy and atomic force microscopy (AFM). The effect of ENO1 silencing was then evaluated by phenotypic and functional experiments to identify the role of ENO1 in adhesion, migration, and invasion, as well as in senescence and apoptosis. The experimental results were then validated in a mouse model.
RESULTS
We observed a significant increase in the roughness of the cell membrane due to ENO1 silencing, a feature associated with an impaired ability to migrate and invade, along with a significant downregulation of proteins involved in cell-cell and cell-matrix adhesion, including alpha v/beta 3 integrin in shENO1 PDA cells. These changes impaired the ability of shENO1 cells to adhere to Collagen I and IV and Fibronectin and caused an increase in RGD-independent adhesion to vitronectin (VN) via urokinase plasminogen activator receptor (uPAR). Binding of uPAR to VN triggers integrin-mediated signals, which result in ERK1-2 and RAC activation, accumulation of ROS, and senescence. In shENO1 cancer cells, the use of an anti-uPAR antibody caused significant reduction of ROS production and senescence. Overall, a decrease of in vitro and in vivo cell migration and invasion of shENO1 PDA cells was observed.
CONCLUSION
These data demonstrate that ENO1 promotes PDA survival, migration, and metastasis through cooperation with integrins and uPAR.
背景
我们之前已经表明，在胰腺导管腺癌（PDA）细胞中，糖酵解酶α-烯醇化酶（ENO1）也作为纤溶酶原受体发挥作用，并促进侵袭和转移形成。此外，PDA细胞中ENO1沉默会诱导氧化应激、衰老并深刻改变PDA细胞代谢。尽管抗ENO1抗体抑制PDA细胞迁移和侵袭，但关于ENO1在调节细胞-细胞和细胞-基质接触中的作用知之甚少。因此，我们研究了ENO1沉默对细胞形态调节、对基质底物的粘附、细胞侵袭性和转移能力的影响。
方法
通过共聚焦显微镜和原子力显微镜（AFM）相结合的方法，研究了在ENO1沉默（shENO1）的PDA细胞中发生的膜和细胞骨架修饰。然后通过表型和功能实验评估ENO1沉默的影响，以确定ENO1在粘附、迁移、侵袭以及衰老和凋亡中的作用。然后在小鼠模型中验证实验结果。
结果
我们观察到由于ENO1沉默，细胞膜粗糙度显著增加，这一特征与迁移和侵袭能力受损相关，同时参与细胞-细胞和细胞-基质粘附的蛋白质显著下调，包括shENO1 PDA细胞中的αv/β3整合素。这些变化损害了shENO1细胞粘附于I型和IV型胶原蛋白以及纤连蛋白的能力，并通过尿激酶型纤溶酶原激活物受体（uPAR）导致对玻连蛋白（VN）的RGD非依赖性粘附增加。uPAR与VN的结合触发整合素介导的信号，导致ERK1-2和RAC激活、活性氧（ROS）积累和衰老。在shENO1癌细胞中，使用抗uPAR抗体可显著降低ROS产生和衰老。总体而言，观察到shENO1 PDA细胞的体外和体内细胞迁移及侵袭减少。
结论
这些数据表明，ENO1通过与整合素和uPAR合作促进PDA的存活、迁移和转移。
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