Mechanism of PAD4-mediated neutrophil extracellular traps promoting proliferation and invasion of thyroid cancer cells
Objective To investigate the mechanism and clinical relevance of peptidyl arginine deiminase 4(PAD4)-mediated neutrophil extracellular traps(NETs)in the proliferation,invasion,and cell cycle of thyroid cancer cells.Methods Peripheral blood of 60 thyroid cancer patients and 60 healthy subjects were collected.Serum PAD4 and citrullinated histone H3(H3cit)levels were detected by ELISA,and the correlations between levels of serum PAD4 and H3cit in thyroid cancer patients and disease progression were analyzed.Neutrophils were extracted from peripheral blood of thyroid cancer patients and healthy subjects,PAD4 mRNA expression levels were detected by qRT-PCR,NETs formation level was detected by ionomycin stimulation and green cyanine staining.Human thyroid cancer cell line CAL-62 cells were co-cultured with NETs and GSK484,and divided into the Control group,the NETs group and the NETs+GSK484 group.CCK-8 assay,Transwell assay,and flow cytometry were used to evaluate the levels of cell proliferation and invasion,and the cell cycle progression,respectively.Results Compared with healthy subjects,the levels of serum PAD4 and H3cit in thyroid cancer patients were increased(P<0.05),and the levels of PAD4 and H3cit were positively correlated(P<0.05).The levels of PAD4 and H3cit in thyroid cancer patients were correlated with TNM stage(P<0.05).Compared with healthy subjects,the expression of PAD4 mRNA and the NETs formation level of peripheral blood neutrophils in thyroid cancer patients increased(P<0.05).Compared with the Control group,the levels of cell proliferation and invasion,and the proportion of S-phase cells increased(P<0.05),while the proportion of G1-phase cells decreased in the NETs group(P<0.05).GSK484 can inhibit the formation of NETs.Compared with the NETs group,the levels of cell proliferation and invasion,and the proportion of S-phase cells decreased(P<0.05),while the proportion of G1-phase cells increased in the NETs+GSK484 group(P<0.05).Conclusion PAD4-mediated NETs promote the proliferation and invasion of thyroid cancer cells and accelerate the progression of thyroid cancer disease by activating cell cycle progression.
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维普
