Department of Pediatrics, Maisonneuve-Rosemont Hospital Research Centre, University of Montreal, Quebec, Canada.
Am J Physiol Regul Integr Comp Physiol. 2011 Mar;300(3):R577-85. doi: 10.1152/ajpregu.00137.2010. Epub 2010 Dec 15.
Cellular migration is a complex process that requires the polymerization of actin filaments to drive cellular extension. Smooth muscle and cancer cell migration has been shown to be affected by coagulation factors, notably the factor VII (FVIIa) and tissue factor (TF) complex. The present studies delineated mediators involved with the process of FVIIa/TF-induced cell migration and utilized a simple, precise, and reproducible, migration assay. Both FVIIa and protease-activated receptor-2 (PAR2)-activating peptide, SLIGRL, increased the migration rate of porcine cerebral microvascular endothelial cells (pCMVECs) overexpressing human TF. Ras homolog gene family member A (RhoA) and cortactin were upregulated during the process; expression of HIF, actin polymerization nuclear diaphanous-related formin-1 and -2 (Dia1, and Dia2) were unaffected. Gene silencing by shRNA to PAR2, RhoA, and cortactin attenuated this gene upregulation and migration induced by FVIIa/TF. Utilizing immunocellular localization, we demonstrate that during FVIIa/TF and PAR2 activation, cortactin molecules translocate from the cytoplasm to the cell periphery and assist in lamellipodia formation of pCMVECs. Overall, we demonstrate a novel regulation and role for cortactin in FVIIa/TF-mediated endothelial cell migration that occurs through a PAR2 and RhoA dependent mechanism.
细胞迁移是一个复杂的过程，需要肌动蛋白丝的聚合来驱动细胞的延伸。已经表明，平滑肌和癌细胞的迁移受到凝血因子的影响，特别是因子 VII（FVIIa）和组织因子（TF）复合物。本研究描绘了与 FVIIa/TF 诱导的细胞迁移过程相关的介质，并利用了一种简单、精确和可重复的迁移测定法。FVIIa 和蛋白酶激活受体 2（PAR2）激活肽 SLIGRL 均增加了过表达人 TF 的猪脑微血管内皮细胞（pCMVEC）的迁移率。在这个过程中，Ras 同源基因家族成员 A（RhoA）和桩蛋白被上调；缺氧诱导因子、肌动蛋白聚合核 diaphanous 相关形态发生因子-1 和 -2（Dia1 和 Dia2）的表达不受影响。通过 shRNA 对 PAR2、RhoA 和桩蛋白的基因沉默减弱了 FVIIa/TF 诱导的这种基因上调和迁移。通过免疫细胞定位，我们证明在 FVIIa/TF 和 PAR2 激活期间，桩蛋白分子从细胞质转移到细胞外周，并协助 pCMVECs 的片状伪足形成。总的来说，我们证明了桩蛋白在 FVIIa/TF 介导的内皮细胞迁移中的新的调节和作用，该作用是通过 PAR2 和 RhoA 依赖的机制发生的。
