Antithrombin controls tumor migration, invasion and angiogenesis by inhibition of enteropeptidase

Abstract

Antithrombin is a key inhibitor of the coagulation cascade, but it may also function as an anti-inflammatory, anti-angiogenic, anti-viral and anti-apoptotic protein. Here, we report a novel function of antithrombin as a modulator of tumor cell migration and invasion. Antithrombin inhibited enteropeptidase on the membrane surface of HT-29, A549 and U-87 MG cells. The inhibitory process required the activation of antithrombin by heparin, and the reactive center loop and the heparin binding domain were essential. Surprisingly, antithrombin non-covalently inhibited enteropeptidase, revealing a novel mechanism of inhibition for this serpin. Moreover, as a consequence of this inhibition, antithrombin was cleaved, resulting in a molecule with anti-angiogenic properties that reduced vessel-like formation of endothelial cells. The addition of antithrombin and heparin to U-87 MG and A549 cells reduced motility in wound healing assays, inhibited the invasion in transwell assays and the degradation of a gelatin matrix mediated by invadopodia. These processes were controlled by enteropeptidase, as demonstrated by RNA interference experiments. Carcinoma cell xenografts in nude mice showed in vivo co-localization of enteropeptidase and antithrombin. Finally, treatment with heparin reduced experimental metastasis induced by HT29 cells in vivo. In conclusion, the inhibition of enteropeptidase by antithrombin may have a double anti-tumor effect through inhibiting a protease involved in metastasis and generating an anti-angiogenic molecule.

Dexamethasone Preconditioning in Cardiac Procedures Reduces Decreased Antithrombin Activity and Is Associated to Beneficial Outcomes: Role of Endothelium

“…Our results show that DX increases SERPINC1 expression and AT promotes migration and angiogenesis in human ECs obtained from larger vessels, whereas DX and AT have no activity on human ECs from microvessels. The same lack of activity for AT, together to an antiangiogenic activity for modified AT was found by other authors in other endothelial cell lines ( Larsson et al, 2001 ; Zhang et al, 2004 , 2005 ; Azhar et al, 2016 ; Luengo-Gil et al, 2016 ).…”

Quantitative proteomic analysis of bile in extrahepatic cholangiocarcinoma patients

“…Antithrombin is known as a protein related with hemostasis, but it has been studied as an antitumor effect through inhibiting a protease involved in metastasis and generating an anti-angiogenic molecule [26]. It was reported as a modulator of tumor cell migration and invasion in gastric cancer [27].…”

Mesothelioma Interactome with 367 Novel Protein-Protein Interactions

“…High expression of GART is observed in cell lines of non-small cell lung cancer; GART and other genes involved in purine biosynthesis such as PPAT, PAICS, PKM2 and ATIC, have been positively correlated with increased proliferation of lung cancer cells (164). One of the novel interactors of GART is a type II transmembrane serine protease that takes part in epithelial-mesenchymal transitions called TMPRSS15, the activity of which is dysregulated in cancer in order to degrade and remodel intercellular junctions and the extracellular matrix (165). Silencing of TMPRSS15 leads to tumor migration and matrix degradation in lung cancer cell lines (165).…”

“…One of the novel interactors of GART is a type II transmembrane serine protease that takes part in epithelial-mesenchymal transitions called TMPRSS15, the activity of which is dysregulated in cancer in order to degrade and remodel intercellular junctions and the extracellular matrix (165). Silencing of TMPRSS15 leads to tumor migration and matrix degradation in lung cancer cell lines (165). NMI, another novel interactor of GART, negatively regulates epithelial-mesenchymal transitions by inhibiting p65 acetylation in the NF-Kβ pathway (166).…”