Neutrophil extracellular traps participate in the development of cancer-associated thrombosis in patients with gastric cancer

BACKGROUND The development of venous thromboembolism(VTE) is associated with high mortality among gastric cancer(GC) patients. Neutrophil extracellular traps(NETs) have been reported to correlate with the prothrombotic state in some diseases, but are rarely reported in GC patients.AIM To investigate the effect of NETs on the development of cancer-associated thrombosis in GC patients.METHODS The levels of NETs in blood and tissue samples of patients were analyzed by ELISA, flow cytometry, and immunofluorescence staining. NET generation and hypercoagulation of platelets and endothelial cells(ECs) in vitro were observed by immunofluorescence staining. NET procoagulant activity(PCA) was determined by fibrin formation and thrombin–antithrombin complex(TAT) assays.Thrombosis in vivo was measured in a murine model induced by flow stenosis in the inferior vena cava(IVC).RESULTS NETs were likely to form in blood and tissue samples of GC patients compared with healthy individuals. In vitro studies showed that GC cells and their conditioned medium, but not gastric mucosal epithelial cells, stimulated NET release from neutrophils. In addition, NETs induced a hypercoagulable state of platelets by upregulating the expression of phosphatidylserine and Pselectin on the cells. Furthermore, NETs stimulated the adhesion of normal platelets on glass surfaces. Similarly, NETs triggered the conversion of ECs to hypercoagulable phenotypes by downregulating the expression of their intercellular tight junctions but upregulating that of tissue factor. Treatment of normal platelets or ECs with NETs augmented the level of plasma fibrin formation and the TAT complex. In the models of IVC stenosis, tumor-bearing mice showed a stronger ability to form thrombi, and NETs abundantly accumulated in the thrombi of tumorbearing mice compared with control mice. Notably, the combination of deoxyribonuclease I,activated protein C, and sivelestat markedly abolished the PCA of NETs.CONCLUSION GC-induced NETs strongly increased the risk of VTE development both in vitro and in vivo. NETs are potential therapeutic targets in the prevention and treatment of VTE in GC patients.