Tissue factor-factor VIIa complex triggers protease activated receptor 2-dependent growth factor release and migration in ovarian cancer

Abstract

Objective Enhanced tissue factor (TF) expression in epithelial ovarian cancer (EOC) is associated with aggressive disease. Our objective was to evaluate the role of the TF-factor VIIa-protease-activated receptor-2 (PAR-2) pathway in human EOC. Methods TCGA RNAseq data from EOC databases were analyzed for PAR expression. Cell and microparticle (MP) associated TF protein expression (Western blot) and MP-associated coagulant activity were determined in human EOC (SKOV-3, OVCAR-3 and CaOV-3) and control cell lines. PAR-1 and PAR-2 protein expression were similarly examined. The PAR dependence of VEGF-A release (ELISA) and chemotactic migration in response to FVIIa and cellular proliferation in response to thrombin was evaluated with small molecule antagonists. Results Relative mRNA expression consistently demonstrated PAR-2>PAR-1≫PAR-3/4 in multiple EOC datasets. Human EOC cell line lysates confirmed expression of TF, PAR-1 and PAR-2 proteins. MPs isolated from EOC cell lines demonstrated markedly enhanced (4–10 fold) TF coagulant activity relative to control cell lines. FVIIa induced a dose-dependent increase in VEGF-A release (2.5-3 fold) from EOC cell lines that was abrogated by the PAR-2 antagonist ENMD-1068. FVIIa treatment of CaOV-3 and OVCAR-3 cells resulted in increased chemotactic migration that was abolished by ENMD-1068. Thrombin induced dose-dependent EOC cell line proliferation was completely reversed by the PAR-1 antagonist vorapaxar. Small molecule antagonists had no effect on these phenotypes without protease present. Conclusions Enhanced activity of the TF-FVIIa-PAR-2 axis may contribute to the EOC progression via PAR-2 dependent signaling that supports an angiogenic and invasive phenotype and local thrombin generation supporting PAR-1 dependent proliferation.

Expression of tissue factor in human cervical carcinoma tissue

“…This may enhance the development of malignant tumors, which suggests that the activation of the coagulation system in certain patients with malignant tumors is closely associated with clinical stage and prognosis ( 13 , 22 ). TFs facilitate the signal transduction of transmembrane proteins in and out of tumor cells and therefore have a greater impact compared with non-transmembrane proteins on the invasion and metastasis of tumor cells ( 23 , 24 ). Previous studies have demonstrated that TF may be activated by forming complexes with coagulation factor VII or VIIa; together with protease activated receptor (PAR)-1 or PAR-2 these complexes may mediate signal transduction within and outside tumor cells ( 23 , 24 ).…”

“…TFs facilitate the signal transduction of transmembrane proteins in and out of tumor cells and therefore have a greater impact compared with non-transmembrane proteins on the invasion and metastasis of tumor cells ( 23 , 24 ). Previous studies have demonstrated that TF may be activated by forming complexes with coagulation factor VII or VIIa; together with protease activated receptor (PAR)-1 or PAR-2 these complexes may mediate signal transduction within and outside tumor cells ( 23 , 24 ). Chanakira et al ( 23 ) revealed that the TF/VIIa complex activates PAR-2 to increase the chemotactic migration of epithelial ovarian tumor cells, and in turn promotes disease progression.…”

A Potent Antagonist of Protease-Activated Receptor 2 That Inhibits Multiple Signaling Functions in Human Cancer Cells

“…They could be helpful molecular tools for teasing out the relative merits of complete versus selective blockade of individual PAR2-linked signaling pathways in cells. In disease settings like cancer, where PAR2 is highly expressed it may be more desirable to inhibit multiple PAR2-dependent signaling associated with metastasis, proliferation, angiogenesis, and other functions promoting tumor development (Dorsam and Gutkind, 2007;Chang et al, 2013;Xie et al, 2015;Chanakira , 2017), whereas in other settings it may be more desirable to dampen some signaling such as that leading to proinflammatory functions while still engaging signaling that leads to beneficial anti-inflammatory actions.…”

Membrane-Anchored Serine Proteases and Protease-Activated Receptor-2–Mediated Signaling: Co-Conspirators in Cancer Progression

“…In addition, a recent survey of PAR family member expression in human tumor samples of various cancer types from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression projects (GTEx) reveals upregulated PAR-2 in 15 different cancer types compared with normal tissues (16). A global transcriptome array analysis of PAR expression in over 1,000 ovarian cancer and normal tissue samples showed that human epithelial ovarian cancers predominantly overexpress PAR-2, followed closely by PAR-1, with minimal detection of PAR-3 and PAR-4 (32). Consistent with this, increased PAR-2 is associated with poor prognosis and decreased progression-free and overall survival in patients with ovarian, cervical, and hepatocellular carcinoma (30,(33)(34)(35).…”