de Oliveira Andreia S, de Almeida Vitor H, Gomes Fausto G, Rezaie Alireza R, Monteiro Robson Q
Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA.
Biochem Biophys Res Commun. 2018 Jan 1;495(1):1300-1304. doi: 10.1016/j.bbrc.2017.11.174. Epub 2017 Nov 28.
Activated Protein C (APC) is a serine-protease that displays antithrombotic and anti-inflammatory properties. In addition, cleavage of protease-activated receptor 1 (PAR1) by APC exerts endothelial cytoprotective actions. The effects of APC on endothelial cells may be reproduced by TR47, a PAR1-based peptide that mimics the novel N-terminus of PAR1 generated upon cleavage at Arg-46 by APC. In this study we demonstrate that wild-type APC and its signaling-proficient mutant, APC-2Cys (which has dramatically reduced anticoagulant activity), display similar inhibitory effects towards the transendothelial migration of A375 human melanoma cells. Consistent with this observation, APC and APC-2Cys significantly reduced the in vivo metastatic potential of the B16F10 murine melanoma cells. TR47 recapitulated the in vitro and in vivo protective profiles of APC and APC-2Cys. Treatment of EA.hy926 endothelial cells with TR47 (20 μM) significantly decreased the A375 cell migration. In addition, treatment of C57/BL6 mice with a single TR47 dose (125 μg/animal) strongly reduced the metastatic burden of B16F10 cells. Together, our results suggest that protection of the endothelial barrier by APC/TR47-mediated signaling pathways might be a valuable therapeutic approach to prevent metastasis.
活化蛋白C（APC）是一种具有抗血栓形成和抗炎特性的丝氨酸蛋白酶。此外，APC对蛋白酶激活受体1（PAR1）的切割发挥内皮细胞保护作用。APC对内皮细胞的作用可由TR47重现，TR47是一种基于PAR1的肽，可模拟APC在Arg-46处切割后产生的PAR1新N端。在本研究中，我们证明野生型APC及其信号传导功能正常的突变体APC-2Cys（其抗凝活性显著降低）对A375人黑色素瘤细胞的跨内皮迁移表现出相似的抑制作用。与该观察结果一致，APC和APC-2Cys显著降低了B16F10小鼠黑色素瘤细胞的体内转移潜能。TR47重现了APC和APC-2Cys的体外和体内保护作用。用TR47（20 μM）处理EA.hy926内皮细胞可显著降低A375细胞迁移。此外，用单一剂量的TR47（125 μg/动物）处理C57/BL6小鼠可强烈降低B16F10细胞的转移负担。总之，我们的结果表明，通过APC/TR47介导的信号通路保护内皮屏障可能是预防转移的一种有价值的治疗方法。
