title: Extracellular vesicles from malignant effusions induce tumor cell migration: inhibitory effect of LMWH tinzaparin.
abstract: Elevated levels of extracellular vesicles (EVs) have been correlated with inflammatory diseases as well as progressive and metastatic cancer. By presenting tissue factor (TF) on their membrane surface, cellular microparticles (MPs) activate both the coagulation system and cell-signaling pathways such as the PAR/ERK pathway. We have shown before that malignant effusions are a rich source of tumor cell-derived EVs. Here, we used EVs from malignant effusions from three different patients after serial low-speed centrifugation steps as recommended by the ISTH (lsEV). Significant migration of human pancreatic carcinoma cells could be induced by lsEVs and was effectively inhibited by pre-incubation with tinzaparin, a low-molecular-weight heparin. Tinzaparin induced tissue factor pathway inhibitor (TFPI) release from tumor cells, and recombinant TFPI inhibited EV-induced tumor cell migration. EVs also induced ERK phosphorylation, whereas inhibitors of PAR2 and ERK suppressed EV-induced tumor cell migration. LsEVs have been characterized by high-resolution flow cytometry and, after elimination of smaller vesicles including exosomes, by further high-speed centrifugation (hsEV). The remaining population consisting primarily of MPs is indeed the main migration-inducing population with tenase activity. Compared to other LMWHs, tinzaparin is suggested to have high potency to induce TFPI release from epithelial cells. The migration-inhibitory effect of TFPI and the interruption of tumor cell migration by inhibitors of PAR2 and ERK suggest that lsEVs induce tumor cell migration by activating the PAR2 signaling pathway. Tinzaparin might inhibit this process at least partly by inducing the release of TFPI from tumor cells, which blocks PAR-activating TF complexes. The clinical relevance of the results is discussed.
authors.affiliation: Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck, Germany.
authors.forename: Hans
authors.initials: H
authors.lastname: Gamperl
authors.affiliation: Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck, Germany.
authors.forename: Corinna
authors.initials: C
authors.lastname: Plattfaut
authors.affiliation: Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck, Germany.
authors.forename: Annika
authors.initials: A
authors.lastname: Freund
authors.affiliation: Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck, Germany.
authors.forename: Tabea
authors.initials: T
authors.lastname: Quecke
authors.affiliation: Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck, Germany.
authors.forename: Friederike
authors.initials: F
authors.lastname: Theophil
authors.affiliation: Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck, Germany. frank.gieseler@uksh.de.
authors.forename: Frank
authors.initials: F
authors.lastname: Gieseler
mesh_terms: D001943:Breast Neoplasms / Q000188:drug therapy / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects* / Q000502:physiology; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000473:pathology; D004730:Endothelium, Vascular / Q000166:cytology; D004847:Epithelial Cells / Q000187:drug effects; D000067128:Extracellular Vesicles / Q000378:metabolism / Q000502:physiology*; D005260:Female; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology*; D006801:Humans; D008074:Lipoproteins / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D016066:Pleural Effusion, Malignant / Q000378:metabolism*; D044464:Receptor, PAR-2 / Q000378:metabolism; D015398:Signal Transduction; D055752:Small Cell Lung Carcinoma / Q000188:drug therapy / Q000473:pathology; D000078222:Tinzaparin
chemical_list: D006495:Heparin, Low-Molecular-Weight; D008074:Lipoproteins; D044464:Receptor, PAR-2; C051928:lipoprotein-associated coagulation inhibitor; D000078222:Tinzaparin
keywords: G-protein-coupled receptors; cell migration; signal peptide/recognition particle