Experimental Neurosurgery, Goethe University Hospital, Frankfurt, Germany.
Neuroscience. 2010 Feb 17;165(4):1312-22. doi: 10.1016/j.neuroscience.2009.11.049. Epub 2009 Dec 1.
Tissue factor (TF) is upregulated in several malignant diseases, including gliomas. Here, we demonstrate pronounced differences in the expression of TF and its interactors factor VII and protease-activated receptor 2 (PAR-2) in nine human glioma cell lines (U87, U251, U343, U373, MZ-18, MZ-54, MZ-256, MZ-304, Hs 683) as detected by RT-PCR and Western blot analysis. Inhibition of TF signaling by a neutralizing monoclonal antibody (mAb TF9-10H10) led to significantly reduced proliferation in high-grade astroglial (MZ-18 and MZ-304) and oligodendroglial (Hs 683) cell lines abundantly expressing TF, but not in U373 cells expressing low amounts of TF. Scratch migration assays and Boyden chamber assays indicated that mAb TF9-10H10 and lentiviral knockdown of TF significantly reduced cell migration and invasion of MZ-18, MZ-304 and Hs 683 cells, both under normoxic and hypoxic conditions. Of note, all three cell lines displayed increased cell migration and invasion under hypoxic conditions (1% O(2)), which was associated with enhanced expression of TF and increased phosphorylation of p44/42 mitogen-activated protein kinase (ERK1/2). Silencing of TF blocked activation of the ERK pathway, induction of TF expression and the potentiating effect of hypoxia on cell migration and invasion. RNA interference against PAR-2 abrogated the autocrine effects of TF on cell proliferation, migration and invasion, indicating that TF signals via PAR-2 in glioma cells. Our results suggest an important role for the TF/FVIIa/PAR-2/ERK axis in tumor growth and invasion of glioma and suggest that TF may be a suitable target for the development of novel therapies against high-grade glioma.
组织因子（TF）在包括神经胶质瘤在内的几种恶性疾病中上调。在这里，我们通过 RT-PCR 和 Western blot 分析证明了 9 个人神经胶质瘤细胞系（U87、U251、U343、U373、MZ-18、MZ-54、MZ-256、MZ-304、Hs 683）中 TF 及其相互作用因子 VII 和蛋白酶激活受体 2（PAR-2）的表达存在明显差异。用中和单克隆抗体（mAb TF9-10H10）抑制 TF 信号导致 TF 表达丰富的高级别星形细胞瘤（MZ-18 和 MZ-304）和少突胶质细胞瘤（Hs 683）细胞系的增殖显著减少，但在 TF 表达量低的 U373 细胞中则没有。划痕迁移实验和 Boyden 室实验表明，mAb TF9-10H10 和慢病毒敲低 TF 显著降低了 MZ-18、MZ-304 和 Hs 683 细胞在常氧和低氧条件下的迁移和侵袭，在低氧条件下（1% O2），所有三种细胞系的迁移和侵袭都增加，这与 TF 的表达增加和丝裂原活化蛋白激酶（ERK1/2）p44/42 的磷酸化增加有关。TF 的沉默阻断了 ERK 途径的激活、TF 表达的诱导以及低氧对细胞迁移和侵袭的增强作用。PAR-2 的 RNA 干扰消除了 TF 对细胞增殖、迁移和侵袭的自分泌作用，表明 TF 通过 PAR-2 在神经胶质瘤细胞中发出信号。我们的研究结果表明，TF/FVIIa/PAR-2/ERK 轴在神经胶质瘤的肿瘤生长和侵袭中具有重要作用，并表明 TF 可能是开发针对高级别神经胶质瘤的新型治疗方法的合适靶点。
