Alpha-enolase (ENO1) controls alpha v/beta 3 integrin expression and regulates pancreatic cancer adhesion, invasion, and metastasis.
Alpha-enolase (ENO1) controls alpha v/beta 3 integrin expression and regulates pancreatic cancer adhesion, invasion, and metastasis.
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DOI：
10.1186/s13045-016-0385-8
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发表时间：
2017-01-13
影响因子：
28.5
通讯作者：
Novelli F
中科院分区：
文献类型：
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作者：
Principe M;Borgoni S;Cascione M;Chattaragada MS;Ferri-Borgogno S;Capello M;Bulfamante S;Chapelle J;Di Modugno F;Defilippi P;Nisticò P;Cappello P;Riganti C;Leporatti S;Novelli F
我们先前已经表明，在胰腺导管腺癌（PDA）细胞中，糖酵解酶α-烯醇化酶（ENO 1）也作为纤溶酶原受体，并促进侵袭和转移形成。此外，在PDA细胞中ENO 1沉默诱导氧化应激、衰老并深刻地改变PDA细胞代谢。尽管抗ENO 1抗体抑制PDA细胞的迁移和侵袭，但关于ENO 1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ENO 1沉默对细胞形态、粘附基质基质、细胞侵袭力和转移能力的调节作用。通过共聚焦显微镜和原子力显微镜（AFM）相结合的方式研究了在ENO 1沉默（shENO 1）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ENO 1沉默的效果，以确定ENO 1在粘附、迁移和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。我们观察到由于ENO 1沉默导致细胞膜粗糙度显著增加，这是与迁移和侵袭能力受损相关的特征，沿着参与细胞-细胞和细胞-基质粘附的蛋白质的显著下调，包括shENO 1 PDA细胞中的α v/β 3整联蛋白。这些变化损害了shENO 1细胞粘附胶原蛋白I和IV和纤连蛋白的能力，并导致通过尿激酶纤溶酶原激活物受体（uPAR）增加与玻连蛋白（VN）的RGD非依赖性粘附。uPAR与VN的结合触发整合素介导的信号，这导致ERK 1 -2和RAC活化、ROS积累和衰老。在shENO 1癌细胞中，使用抗uPAR抗体导致ROS产生和衰老显著减少。总之，观察到shENO 1 PDA细胞的体外和体内细胞迁移和侵袭减少。这些数据表明，ENO 1通过与整合素和uPAR的合作促进PDA存活、迁移和转移。本文的在线版本（doi：10.1186/s13045-016-0385-8）包含补充材料，可供授权用户使用。
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. 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. 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. These data demonstrate that ENO1 promotes PDA survival, migration, and metastasis through cooperation with integrins and uPAR. The online version of this article (doi:10.1186/s13045-016-0385-8) contains supplementary material, which is available to authorized users.