Expression of IL-17A in esophageal adenocarcinoma and its regulatory effect on NF-κB/COX-2 signaling axis
Objective To explore the expression of interleukin 17A(IL-17A)in esophageal adenocarcinoma and its role and mecha-nism in the regulation of cyclooxygenase-2(COX-2)protein through the nuclear transcription factor κB(NF-κB)signaling pathway.Methods The specimens of esophageal adenocarcinoma,paracancerous tissues and serum were collected from ten patients with esopha-geal adenocarcinoma.At the same time,normal esophageal epithelial tissues and serum were collected from ten healthy subjects undergoing gastroscopy.Expressions of IL-17A and COX-2 proteins were detected in tissues by immunohistochemical assay.IL-17A mRNA and retinoic acid receptor related orphan c(RORc)mRNA,a specific transcription factor of Th17 cells,were detected in cancerous tissues and paracancerous tissues by real time-PCR method.The serum levels of IL-17A were measured in esophageal adeno-carcinoma patients and healthy controls by ELISA method.Human esophageal adenocarcinoma OE19 cells in logarithmic growth phase were divided into control group,IL-17A(100 ng/mL)group and IL-17A(100 ng/mL)+PDTC(100 μmol/L,NF-κB signaling pathway inhibitor pyrrolidine di thiocarbamate)group,and the expressions of p50,p65,p-IκB-α,and COX-2 proteins were detected by Western blotting in OE19 cells.Results The number of IL-17A-positive cells was higher in cancer tissues than in paracancerous tissues.The levels of IL-17A mRNA and RORc mRNA were increased in esophageal adenocarcinoma tissues compared to paracancerous tissues(P＜0.05).Serum level of IL-17A was significantly higher in patients with esophageal adenocarcinoma than in healthy controls(P＜0.05).Compared with normal esophageal epithelial tissues,there was a larger and deeper positive staining area of COX-2 protein in esophageal adenocarcinoma tissues.The expressions of p-IκB-α and COX-2 proteins in OE19 cells in IL-17A group was significantly increased compared with control group(P＜0.05),and the expressions of p-IKB-α and COX-2 proteins in OE19 cells in IL-17A+PDTC group were significantly decreased compared with IL-17A group(P＜0.05).Conclusion IL-17A expression is increased in both tissues and serum of patients with esophageal adenocarcinoma.IL-17A could upregulate the expression of COX-2 in OE19 cells by activating the NF-κB signaling pathway.
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