Inhibition of invasion and experimental metastasis of murine melanoma cells by human soluble thrombomodulin
Inhibition of invasion and experimental metastasis of murine melanoma cells by human soluble thrombomodulin
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DOI：
10.1016/s0304-3835(00)00617-0
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发表时间：
2000-12-20
期刊：
影响因子：
9.7
通讯作者：
Ishii, H
中科院分区：
文献类型：
--
作者：
Hosaka, Y;Higuchi, T;Ishii, H
血栓调节蛋白（TM）是一种表达于内皮细胞表面的抗凝分子，可溶性TM抗原存在于人血浆和尿液中，代表细胞表面TM的有限蛋白水解裂解产物。近年来研究表明，TM在多种肿瘤细胞表面也有表达，其表达水平与肿瘤的恶性程度呈负相关。我们研究了从人尿中分离的可溶性TM（uTM）对小鼠黑色素瘤细胞（B16 F10细胞）通过重建基底膜（Matrigel）和小鼠实验性肺转移模型的侵袭和转移的影响。与不含uTM的对照Matrigel相比，在10至1000 ng/ml uTM范围内，用uTM重建的Matrigel以剂量依赖性方式抑制B16 F10细胞的侵袭。uTM的抑制作用在过量水蛭素（凝血酶蛋白水解活性的抑制剂）的存在下没有改变，但在抗人TM IgG的存在下被废除。相对于不含凝血酶的对照Matrigel，用凝血酶（1 NIH单位/ml）重建的Matrigel使细胞的侵袭水平提高了1.5倍。凝血酶增强的B16 F10细胞的侵袭通过向Matrigel中加入水蛭素（10单位/ml）或uTM（100 ng/ml）来抑制。用水蛭素（10单位/ml）和uTM（100 ng/ml）重构的基质胶另外加速水蛭素或uTM对凝血酶增强的B16 F10细胞侵袭的抑制活性。此外，通过每天一次注射uTM，直到uTM与细胞共注射后2天，静脉内注射B16 F10细胞的小鼠的肺中形成的转移性集落显著减少。这些结果表明，与uTM重组的Matrigel通过凝血酶非依赖性机制抑制B16 F10细胞的体外侵袭，并且注射uTM抑制细胞在小鼠中的实验性肺转移。(C)2000爱思唯尔科学爱尔兰有限公司保留所有权利。
Thrombomodulin (TM) is an anticoagulant molecule expressed on the endothelial cell surface and soluble TM antigen, which is present in human plasma and urine, represents the products of limited proteolytic cleavage of cell-surface TM. Recently, it was demonstrated that TM is also expressed on the surface of several tumor cells and the expression level of TM negatively correlated with malignancy in cancer. We investigated the effect of soluble TM isolated from human urine (uTM) on the invasion and metastasis of murine melanoma cells (B16F10 cells) through a reconstituted basement membrane (Matrigel) and in a murine model of experimental lung metastasis. Matrigel reconstituted with uTM inhibited the invasion of B16F10 cells in a dose-dependent manner in a range from 10 to 1000 ng/ml uTM as compared with the control Matrigel without uTM. The inhibitory action of uTM was not altered in the presence of an excess amount of hirudin, an inhibitor of thrombin proteolytic activity, but abolished in the presence of anti-human TM IgG. Matrigel reconstituted with thrombin (1 NIH unit/ml) enhanced the invasion level of cells by 1.5-fold relative to the control Matrigel without thrombin. The thrombin-enhanced invasion of B16F10 cells was repressed by addition of hirudin (10 units/ml) or uTM (100 ng/ml) into the Matrigel. Matrigel reconstituted with hirudin (10 units/ml) and uTM (100 ng/ml) additionally accelerated the inhibitory activity of hirudin or uTM on the thrombin-enhanced invasion of B16F10 cells. Moreover, metastatic colonies formed in the lungs of mice injected intravenously with B16F10 cells were significantly reduced by injection of uTM once a day up to 2 days after co-injection of uTM with the cells. These results suggested that Matrigel reconstituted with uTM inhibited the invasion of B16F10 cells in vitro through a thrombin-independent mechanism and the injection of uTM suppressed experimental lung metastasis of the cells in mice. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.