Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1524, Prédio I, sala 428 05508-000, São Paulo, SP, Brazil.
Centro de Facilidades para a Pesquisa (CEFAP), Instituto de Ciência Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1730, Prédio IV, 05508-000, São Paulo, SP, Brazil.
Exp Cell Res. 2018 Feb 15;363(2):271-282. doi: 10.1016/j.yexcr.2018.01.017. Epub 2018 Jan 31.
Extracellular matrix (ECM) serves as a reservoir for biologically active factors, such as growth factors and proteases that influence the tumor cell behavior. ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs) is a secreted protease that has the ability to modify the ECM during physiological and pathological processes. Here, we analyzed the role played by ADAMTS-1 regulating HGF and TGF-β1 activities in the high-grade fibrosarcoma cell line (HT1080). We generated HT1080 and HEK293T cells overexpressing ADAMTS-1. HT1080 cells overexpressing ADAMTS-1 (HT1080-MPA) exhibited a significant decrease in cell proliferation and migration velocity, both in presence of HGF. We obtained similar results with ADAMTS-1-enriched conditioned medium from other cell type. However, ADAMTS-1 overexpression failed to affect TGF-β1 activity associated with HT1080 cell proliferation and migration velocity. Immunoblotting showed that ADAMTS-1 overexpression disturbs c-Met activation upon HGF stimulation. Downstream ERK1/2 and FAK signaling pathways are also influenced by this protease. Additionally, ADAMTS-1 decreased the size of the fibrosarcospheres, both under normal conditions and in the presence of HGF. Likewise, in presence of HGF, ADAMTS-1 overexpression in HT1080 disrupted microtumors formation in vivo. These microtumors, including individual cells, presented characteristics of non-invasive lesions (rounded morphology). Our results suggest that ADAMTS-1 is involved in regulating HGF-related functions on fibrosarcoma cells. This protease may then represent an endogenous mechanism in controlling the bioavailability of different growth factors that have a direct influence on tumor cell behavior.
细胞外基质（ECM）充当生物活性因子的储库，例如影响肿瘤细胞行为的生长因子和蛋白酶。ADAMTS-1（解整合素和金属蛋白酶与血栓反应蛋白基序）是一种分泌型蛋白酶，具有在生理和病理过程中修饰 ECM 的能力。在这里，我们分析了 ADAMTS-1 调节 HGF 和 TGF-β1 活性在高级纤维肉瘤细胞系（HT1080）中的作用。我们生成了过表达 ADAMTS-1 的 HT1080 和 HEK293T 细胞。过表达 ADAMTS-1 的 HT1080 细胞（HT1080-MPA）在存在 HGF 的情况下表现出细胞增殖和迁移速度的显着降低。我们从其他细胞类型获得了类似的结果与富含 ADAMTS-1 的条件培养基。然而，ADAMTS-1 过表达未能影响与 HT1080 细胞增殖和迁移速度相关的 TGF-β1 活性。免疫印迹显示 ADAMTS-1 过表达扰乱了 HGF 刺激后的 c-Met 激活。ERK1/2 和 FAK 信号通路的下游也受到这种蛋白酶的影响。此外，ADAMTS-1 减小了纤维肉瘤球体的大小，无论是在正常条件下还是在 HGF 存在下。同样，在 HGF 存在下，HT1080 中的 ADAMTS-1 过表达破坏了体内微肿瘤的形成。这些微肿瘤，包括单个细胞，表现出非侵袭性病变的特征（圆形形态）。我们的结果表明 ADAMTS-1 参与调节纤维肉瘤细胞与 HGF 相关的功能。这种蛋白酶可能代表一种内源性机制，用于控制对肿瘤细胞行为有直接影响的不同生长因子的生物利用度。
