Vasodilator-Stimulated Phosphoprotein (VASP) depletion from breast cancer MDA-MB-231 cells inhibits tumor spheroid invasion through downregulation of Migfilin, β-catenin and urokinase-plasminogen activator (uPA).
Vasodilator-Stimulated Phosphoprotein (VASP) depletion from breast cancer MDA-MB-231 cells inhibits tumor spheroid invasion through downregulation of Migfilin, β-catenin and urokinase-plasminogen activator (uPA).
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DOI：
10.1016/j.yexcr.2017.02.019
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发表时间：
2017-03-15
影响因子：
3.7
通讯作者：
Stylianopoulos T
中科院分区：
文献类型：
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作者：
Gkretsi V;Stylianou A;Stylianopoulos T
癌细胞的标志是它们侵入周围组织并形成转移的能力。细胞-细胞外基质（ECM）-粘附蛋白在转移中至关重要，将肿瘤ECM与肌动蛋白细胞骨架连接，从而使细胞能够响应机械信号。血管舒张刺激磷蛋白（VASP）是一种肌动蛋白聚合调节剂，与细胞-ECM粘附蛋白Migfilin相互作用，调节细胞迁移。我们比较了VASP在MCF-7和MDA-MB-231乳腺癌（BC）细胞中的表达，发现更具侵袭性的MDA-MB-231细胞过表达VASP。然后，我们利用三维（3D）的方法来研究转移MDA-MB-231细胞使用的系统，认为ECM施加的机械力。我们制备了浓度增加的3D I型胶原凝胶，通过原子力显微镜对其进行成像，并在存在或不存在VASP的情况下将其用于包埋细胞或肿瘤球体。我们首次发现，VASP沉默下调Migfilin，β-catenin和尿激酶纤溶酶原激活剂在2D和3D，这表明一个矩阵独立的机制。缺乏VASP的肿瘤球状体表现出受损的侵袭，表明VASP参与转移，Kaplan-Meier检验证实了这一点，Kaplan-Meier检验显示VASP高表达与淋巴结阳性BC患者的无缓解生存率差相关。因此，VASP可能是一种新的BC转移生物标志物。
A hallmark of cancer cells is their ability to invade surrounding tissues and form metastases. Cell-extracellular matrix (ECM)-adhesion proteins are crucial in metastasis, connecting tumor ECM with actin cytoskeleton thus enabling cells to respond to mechanical cues. Vasodilator-stimulated phosphoprotein (VASP) is an actin-polymerization regulator which interacts with cell-ECM adhesion protein Migfilin, and regulates cell migration. We compared VASP expression in MCF-7 and MDA-MB-231 breast cancer (BC) cells and found that more invasive MDA-MB-231 cells overexpress VASP. We then utilized a 3-dimensional (3D) approach to study metastasis in MDA-MB-231 cells using a system that considers mechanical forces exerted by the ECM. We prepared 3D collagen I gels of increasing concentration, imaged them by atomic force microscopy, and used them to either embed cells or tumor spheroids, in the presence or absence of VASP. We show, for the first time, that VASP silencing downregulated Migfilin, β-catenin and urokinase plasminogen activator both in 2D and 3D, suggesting a matrix-independent mechanism. Tumor spheroids lacking VASP demonstrated impaired invasion, indicating VASP’s involvement in metastasis, which was corroborated by Kaplan-Meier plotter showing high VASP expression to be associated with poor remission-free survival in lymph node-positive BC patients. Hence, VASP may be a novel BC metastasis biomarker.