Neutrophil extracellular traps regulating the development and prognosis of oral squamous cell carcinomas

Abstract

Objective To investigate the expression and role of neutrophil extracellular traps(NETs) in oral squamous cell carcinoma(OSCC). Methods Immunofluorescence and immunohistochemistry were used to detect the expression of NETs in OSCC. The clinicopathological characteristics and prognosis of patients with different NETs expression levels were analyzed. Density gradient centrifugation was used to isolate neutrophils from human peripheral blood. NETs were induced by phorbol 12-myristate 13-acetate(PMA) and co-cultured with OSCC cells. CCK-8 assay was used to detect changes in OSCC cell proliferation ability. OSCC cell migration ability was detected by Transwell cell migration assay and cell scratch assay. Western blotting(WB) assay was used to detect the effect of NETs on the index of epithelial-mesenchymal transition(EMT). Results NETs’ expression in OSCC was higher than that in normal tissues(P<0.001). The prognosis of patients with high NETs expression was worse than that of patients with low NETs expression(P<0.05). The expression level of NETs was correlated with the clinical grade, invasion and recurrence degree of OSCC patients(P<0.05). NETs promoted the proliferation and migration of Cal27 and HN6 cells(P<0.05), and inhibited the protein expression level of epithelial marker and promoted the protein expression level of mesenchymal markers(P<0.05), which could be reversed by the NETs inhibitor DNaseⅠ. Conclusion NETs are expressed at high levels in OSCC. NETs can promote the proliferation and migration of OSCC cells by regulating epithelial-mesenchymal transition and can affect the prognosis of OSCC patients.

Keywords

• |neutrophil extracellular traps|oral squamous cell carcinoma|epithelial-mesenchymal transition