Platelet-Derived TGF-β Induces Tissue Factor Expression via the Smad3 Pathway in Osteosarcoma Cells
Platelet-Derived TGF-β Induces Tissue Factor Expression via the Smad3 Pathway in Osteosarcoma Cells
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DOI：
10.1002/jbmr.3537
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发表时间：
2018-11-01
影响因子：
6.2
通讯作者：
Haro, Hirotaka
中科院分区：
文献类型：
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作者：
Saito, Masanori;Ichikawa, Jiro;Haro, Hirotaka
在过去的三十年里，骨肉瘤的预后一直没有改变;肺转移患者的预后仍然很差，迫切需要开发新的治疗方法。我们以前的研究表明，侵袭性骨肉瘤细胞表达更多的组织因子（TF），并表现出增强的外源性途径的能力。此外，抗凝血剂低分子量肝素可抑制肿瘤生长。然而，TF调节的分子机制仍不清楚。在这里，我们报告说，转化生长因子-β（TGF-β）上调TF，这可以通过激活血小板发生。发现TF在骨肉瘤细胞表面表达，其介导Xa和凝血酶的产生。在几种骨肉瘤细胞中观察到TGF-β诱导TF，尤其是在MG 63细胞中。通过TGF-β表达的TF和通过TF的外源性途径活性均迅速增加。该反应被TGF-β I型受体抑制剂和TGF-β中和抗体抑制。虽然发现TGF-β磷酸化Smad 2和Smad 3，但它们的作用明显不同。令人惊讶的是，Smad 2敲低没有导致抑制作用，而Smad 3敲低完全抑制TGF-β诱导的TF表达。接下来，数据表明血小板是TGF-β的来源。我们证实，凝血酶激活的血小板和骨肉瘤细胞可以释放TGF-β，血小板衍生的TGF-β可以诱导TF表达。这些过程也被TGF-β I型受体抑制剂和Smad 3敲低抑制。此外，还在来自骨肉瘤患者的活检样品中检测到CD 42 b、TF、TGF-β、Smad 2/3和p-Smad 2/3。总的来说，这些发现表明，骨肉瘤细胞和血小板之间的相互作用，通过凝血酶和TGF-β，导致一个连续的周期，抗血小板或抗TGF-β治疗可能是一个有前途的工具，用于疾病治疗。(c)2018年美国骨与矿物质研究学会。
Over the last three decades, the prognosis of osteosarcoma has remained unchanged; the prognosis for patients with lung metastasis is still poor, and the development of new treatments is urgently required. We previously showed that aggressive osteosarcoma cells express more tissue factor (TF) and demonstrate enhanced extrinsic pathway capacity. Furthermore, tumor growth can be suppressed with the anticoagulant low molecular weight heparin. However, the molecular mechanisms underlying TF regulation are still unclear. Here, we report that transforming growth factor-beta (TGF-beta) upregulates TF, which can occur via activated platelets. TF was found to be expressed on osteosarcoma cell surfaces, which mediated the production of Xa and thrombin. TF induction by TGF-beta was observed in several osteosarcoma cells, and especially in MG 63 cells. Both TF expression by TGF-beta and extrinsic pathway activity through TF were rapidly increased. This reaction was inhibited by a TGF-beta type I receptor inhibitor and TGF-beta neutralizing antibody. Although TGF-beta was found to phosphorylate both Smad2 and Smad3, their roles were markedly disparate. Surprisingly, Smad2 knockdown resulted in no inhibitory effect, whereas Smad3 knockdown completely suppressed TGF-beta-induced TF expression. Next, data suggested that platelets were the source of TGF-beta. We confirmed that thrombin-activated platelets and osteosarcoma cells could release TGF-beta, and that platelet-derived TGF-beta could induce TF expression. These processes were also inhibited by a TGF-beta type I receptor inhibitor and Smad3 knockdown. Moreover, CD42b, TF, TGF-beta, Smad2/3, and p-Smad2/3 were also detected in a biopsy sample from an osteosarcoma patient. Collectively, these finding suggested that the interaction between osteosarcoma cells and platelets, via thrombin and TGF-beta, results in a continuous cycle, and that anti-platelet or anti-TGF-beta therapy could be a promising tool for disease treatment. (c) 2018 American Society for Bone and Mineral Research.