Department of Orthopaedic, China Medical University Hospital, Taichung, Taiwan.
J Cell Physiol. 2010 Jun;223(3):737-45. doi: 10.1002/jcp.22083.
Thrombin is a multifunctional protease that can activate hemostasis and coagulation through the cleavage of fibrinogen to form fibrin clots. Thrombin also plays a crucial role in migration and metastasis of human cancer cells. However, the effect of thrombin on migration activity in human chondrosarcoma cells is mostly unknown. Here, we found that thrombin increased the migration and expression of matrix metalloproteinase (MMP)-2 and MMP-13 in human chondrosarcoma cells (JJ012 and SW1353 cells). By using pharmacological inhibitors or activators or genetic inhibition by the protease-activated receptor (PAR), we found that the PAR1 and PAR4 receptor but not PAR3 receptor are involved in thrombin-mediated cell migration and MMPs expression. Thrombin-mediated migration and MMPs up-regulation was attenuated by phospholipase C (PLC), protein kinase C, and c-Src inhibitor. Activations of PLCbeta, PKCalpha, c-Src, and NF-kappaB pathways after thrombin treatment was demonstrated, and thrombin-induced MMPs expression and migration activity was inhibited by the specific inhibitors and mutants of PLC, PKC, c-Src, and NF-kappaB cascades. Taken together, our results indicated that thrombin enhances the migration of chondrosarcoma cells by increasing MMP-2 and MMP-13 expression through the PAR/PLC/PKCalpha/c-Src/NF-kappaB signal transduction pathway.
凝血酶是一种多功能蛋白酶，可通过裂解纤维蛋白原形成纤维蛋白凝块来激活止血和凝血。凝血酶还在人类癌细胞的迁移和转移中起着关键作用。然而，凝血酶对人软骨肉瘤细胞迁移活性的影响在很大程度上尚不清楚。在这里，我们发现凝血酶增加了人软骨肉瘤细胞（JJ012 和 SW1353 细胞）中的迁移和基质金属蛋白酶（MMP）-2 和 MMP-13 的表达。通过使用药理学抑制剂或激活剂或蛋白酶激活受体（PAR）的基因抑制，我们发现 PAR1 和 PAR4 受体而非 PAR3 受体参与了凝血酶介导的细胞迁移和 MMPs 表达。凝血酶介导的迁移和 MMPs 的上调可被磷脂酶 C（PLC）、蛋白激酶 C 和 c-Src 抑制剂减弱。证明了凝血酶处理后 PLCβ、PKCalpha、c-Src 和 NF-κB 途径的激活，并且 PLC、PKC、c-Src 和 NF-κB 级联的特异性抑制剂和突变体抑制了凝血酶诱导的 MMPs 表达和迁移活性。总之，我们的结果表明，凝血酶通过 PAR/PLC/PKCalpha/c-Src/NF-κB 信号转导通路增加 MMP-2 和 MMP-13 的表达来增强软骨肉瘤细胞的迁移。
