Expression Of IL-33 And Its Receptor ST2 In Esophageal Adenocarcinoma And Their Effects On Proliferation,Migration And Invasion Of Esophageal Adenocarcinoma Cells

Objective: To investigate the expression of IL-33 and its specific receptor ST2 in esophageal adenocarcinoma and their effects on proliferation,migration and invasion of esophageal adenocarcinoma cells,and to explore whether the intervention of IL-33/ST2 pathway may provide a new strategy for the prevention and treatment of esophageal adenocarcinoma.Methods: 1.Immunohistochemistry(IHC),Real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot(WB)were used to detect the expression of IL-33 and ST2 in the esophageal adenocarcinoma and normal esophageal epithelium,also in the esophageal adenocarcinoma cell line OE33 and the normal esophageal epithelial cell line HEEC:(1)The expression of IL-33 and ST2 in esophageal adenocarcinoma and normal esophageal epithelial tissue was compared by immunohistochemical staining and grading according to the intensity and range of tissue staining.(2)Total RNA was extracted from esophageal adenocarcinoma,normal esophageal epithelium and OE33,HEEC cells by TRIzol method.The expression of IL-33 was detected by RTqPCR.(3)Proteins were extracted from esophageal adenocarcinoma,normal esophageal epithelium and OE33,HEEC cells.The expressions of IL-33 and ST2 were detected by Western blot.2.The relationship between the expression of IL-33 and clinical pathology and prognosis of patients:(1)The correlation between the expression of IL-33 and tumor parameters,such as age,sex,pathological stage and grading(such as tumor size,differentiation,depth of invasion,lymph node metastasis and distant metastasis),was analyzed by tissue microarray including esophageal adenocarcinoma and normal esophageal epithelial tissue adjacent to cancer.(2)The relationship between the expression of IL-33 and prognosis was analyzed by Kaplan-meier survival curve.3.The effects of exogenous IL-33 on proliferation,migration and invasion of esophageal adenocarcinoma cells were investigated by cell function experiments:(1)CCK-8 test was used to detect the effect of IL-33 on the growth and proliferation of esophageal adenocarcinoma cells;(2)Wound healing test was used to observe the effect of IL-33 on the migration ability of esophageal adenocarcinoma cells;(3)Transwell test was used to observe the effect of IL-33 on the invasion ability of esophageal adenocarcinoma cells;(4)After blocking the IL-33/ST2 pathway with sST2(ST2 neutralizing antibody),the effects on the above experimental results were observed;(5)Western blot was used to detect the effect of IL-33 on the epithelial-mesenchymal transition(EMT)of esophageal adenocarcinoma cells.Results: 1.Immunohistochemical results showed that the expression of IL-33 and ST2 in esophageal adenocarcinoma was significantly higher than that in normal esophageal epithelium.IL-33 was mainly expressed in the cytoplasm of tumor cells and in the nucleus of normal esophageal epithelium cells.ST2 was expressed in the cytoplasm and membrane of both esophageal adenocarcinoma and normal tissues,and its expression was higher in adenocarcinoma tissues.RTqPCR results showed that the expression of IL-33 mRNA in esophageal adenocarcinoma was higher than that in normal esophageal epithelium,and the expression of IL-33 mRNA in esophageal adenocarcinoma cell line OE33 was higher than that in normal esophageal epithelium cell line HEEC.Western blot results showed that the protein level of IL-33 and ST2 in esophageal adenocarcinoma and esophageal adenocarcinoma cells was significantly higher than that in normal esophageal epithelium and normal esophageal epithelium cells.2.Tissue microarray analysis of 64 cases of esophageal adenocarcinoma and adjacent tissues showed that the high expression of IL-33 was positively correlated with the grade of T1-2 and the degree of moderate to high differentiation of esophageal adenocarcinoma.Survival curve analysis showed that the expression of IL-33 was not correlated with the cumulative survival rate of patients.3.In vivo experiments showed that exogenous IL-33 promoted the proliferation,migration and invasion of esophageal adenocarcinoma cells OE33,which was blocked by sST2,and IL-33 induced EMT of OE33.Conclusion: IL-33 and its receptor ST2 are highly expressed in esophageal adenocarcinoma.IL-33/ST2 pathway can promotes the proliferation,migration and invasion of esophageal adenocarcinoma cells and participate in the occurrence and development of tumors.