Abstract
Background
Hypercoagulability is a major cancer-associated complication linked to poor patient prognosis. The production of neutrophil extracellular traps (NETs) is increasingly found to be linked with the development and metastasis of cancer, as well as with thrombi formation in cancer patients. We hypothesized that the neutrophil NET release may be triggered by specific cytokines in oral squamous cell carcinoma (OSCC) patients, thereby predisposing them to a hypercoagulable state. Moreover, we have evaluated the interaction between NETs and endothelial cells (ECs).
Methods
NET procoagulant activity was assessed based on fibrin and purified coagulation complex production assays, as well as by measuring coagulation time (CT). We further used confocal microscopy to quantify the exposure of phosphatidylserine (PS), fibrin strands, and cell FVa/Xa binding.
Results
OSCC patients with stage III/IV exhibited elevated plasma NET levels compared to stage I/II or CTR (all P < 0.05). Neutrophils from OSCC patients are predisposed to amplified NET release compared to those from CTR. Furthermore, depleting IL-8, IL-6, and TNF-α led to a reduction in NET release in the plasma. OSCC NETs increased thrombin and fibrin generation and decreased CT significantly (P < 0.05). When NETs were isolated and used to treat ECs, these cells exhibited disrupted morphology by retracting from their cell–cell junctions and convert to a procoagulant phenotype. These effects could be attenuated by approximately 70% using DNase I.
Conclusions
Our findings are consistent with a model wherein OSCC drives a systemic inflammatory state, which, in turn, drives neutrophils to prime and release NETs, which drive the development of a hypercoagulable state. Intervening in this process may be a viable means of disrupting these undesirable coagulation dynamics in stage III/IV OSCC patients.
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Acknowledgements
We thank Yue Cao and Lu Wang for the sample collection.
Funding
This study was supported by the National Natural Science Foundation of China (81470301, 81670128, and 81873433) and Harbin Science and Technology Bureau (2005AA9CS116-3).
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Study concept and design (BL, JS, and TH); data acquisition, clinical data and sample collection, statistical analysis (BL and YL); data interpretation and manuscript drafting (BL, JS, TH); critical revision of the manuscript (BL, VN, YL, YZ, TL, JS, and TH); YZ, CZ, TL, CW, TH, ZD, VN, JS, and TH provided valuable advice for this study. All authors have read and approved the final manuscript.
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Li, B., Liu, Y., Hu, T. et al. Neutrophil extracellular traps enhance procoagulant activity in patients with oral squamous cell carcinoma. J Cancer Res Clin Oncol 145, 1695–1707 (2019). https://doi.org/10.1007/s00432-019-02922-2
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DOI: https://doi.org/10.1007/s00432-019-02922-2