Department of Internal Medicine, Division of Hematology/Oncology, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America.
Mol Med. 2012 Jul 18;18(1):771-9. doi: 10.2119/molmed.2011.00416.
This study was performed to determine whether murine alternatively spliced tissue factor (masTF) acts analogously to human alternatively spliced tissue factor (hasTF) in promoting neovascularization via integrin ligation. Immunohistochemical evaluation of a spontaneous murine pancreatic ductal adenocarcinoma model revealed increased levels of masTF and murine full-length tissue factor (mflTF) in tumor lesions compared with benign pancreas; furthermore, masTF colocalized with mflTF in spontaneous aortic plaques of Ldlr(-/-) mice, indicating that masTF is likely involved in atherogenesis and tumorigenesis. Recombinant masTF was used to perform in vitro and ex vivo studies examining its integrin-mediated biologic activity. Murine endothelial cells (ECs) rapidly adhered to masTF in a β3-dependent fashion. Using adult and embryonic murine ECs, masTF potentiated cell migration in transwell assays. Scratch assays were performed using murine and primary human ECs; the effects of masTF and hasTF were comparable in murine ECs, but in human ECs, the effects of hasTF were more pronounced. In aortic sprouting assays, the potency of masTF-triggered vessel growth was undistinguishable from that observed with hasTF. The proangiogenic effects of masTF were found to be Ccl2-mediated, yet independent of vascular endothelial growth factor. In murine ECs, masTF and hasTF upregulated genes involved in inflammatory responses; murine and human ECs stimulated with masTF and hasTF exhibited increased interaction with murine monocytic cells under orbital shear. We propose that masTF is a functional homolog of hasTF, exerting some of its key effects via β3 integrins. Our findings have implications for the development of murine models to examine the interplay between blood coagulation, atherosclerosis and cancer.
本研究旨在确定小鼠选择性剪接组织因子（masTF）是否通过整合素结合类似于人选择性剪接组织因子（hasTF）促进血管新生。对自发性小鼠胰腺导管腺癌模型的免疫组织化学评估显示，与良性胰腺相比，肿瘤病变中 masTF 和小鼠全长组织因子（mflTF）的水平升高；此外，masTF 与 Ldlr(-/-) 小鼠自发性主动脉斑块中的 mflTF 共定位，表明 masTF 可能参与动脉粥样硬化和肿瘤发生。使用重组 masTF 进行体外和离体研究，以检查其整合素介导的生物学活性。小鼠内皮细胞（ECs）以 β3 依赖性方式快速附着于 masTF。使用成年和胚胎期小鼠 ECs，masTF 增强了 Transwell 测定中的细胞迁移。使用小鼠和原代人 ECs 进行划痕实验；masTF 和 hasTF 在小鼠 ECs 中的作用相当，但在人 ECs 中，hasTF 的作用更为明显。在主动脉发芽测定中，masTF 触发的血管生长的效力与观察到的 hasTF 没有区别。masTF 的促血管生成作用被发现是 Ccl2 介导的，但独立于血管内皮生长因子。在小鼠 ECs 中，masTF 和 hasTF 上调参与炎症反应的基因；用 masTF 和 hasTF 刺激的小鼠和人 ECs 在轨道剪切下与小鼠单核细胞的相互作用增加。我们提出，masTF 是 hasTF 的功能同源物，通过β3 整合素发挥其一些关键作用。我们的发现对开发研究血液凝固、动脉粥样硬化和癌症之间相互作用的小鼠模型具有重要意义。
