Expression And Significance Of GSDMD In Renal Clear Cell Carcinoma

Objective:To explore the expression of Gasdermins D(GSDMD)in renal clear cell carcinoma and its effect on the proliferation,apoptosis,invasion and migration of renal clear cell carcinoma.Method:The TCGA database was used to compare the expression of GSDMD in tumor and normal tissues,analyze its correlation with cancer stage and total survival time,and establish receiver operating characteristic curve(ROC curve).The results were confirmed by immunohistochemical staining of 65 clinical samples and PCR and WB of three cell lines.TCGA database was used to screen the differentially expressed genes(EDGs)of GSDMD,and Gene MANIA and STRING were used to construct the protein interaction network(PPI)of GSDMD,and R software was used to conduct GO and KEGG enrichment analysis of related genes.Use lentivirus with high expression and knockdown GSDMD to transfect 769-P cells to construct stable up-regulated and down-regulated transfected cell lines,and then MTT to detect the proliferation of the two groups of transfected cells,flow cytometry to detect the apoptosis of the two groups of cells,and Transwell to detect migration and invasion.Results:Among the 33 kinds of cancer,15 kinds of cancer,including renal clear cell carcinoma,have increased expression of GSDMD.The expression of GSDMD in renal clear cell carcinoma is significantly related to the stage and total survival of patients.The tumor stage and total survival of patients with high expression of GSDMD are worse.Through ROC curve analysis,GSDMD has certain significance in the diagnosis of renal clear cell carcinoma(AUC=0.855).Through the immunohistochemical examination of 65 patients with renal clear cell carcinoma(RCC),it was verified that the increased expression of GSDMD in RCC was significantly correlated with the stage,grade,sex and smoking of the patients(P<0.05),but not with age,tumor location,tumor size,high blood pressure,clinical symptoms and BMI level(P>0.05).In addition,patients with high expression of GSDMD have poorer disease-free survival,and multivariate COX regression analysis shows that GSDMD is an independent risk factor affecting disease-free survival of renal clear cell carcinoma.After that,two groups of transfected cells with high expression and knockdown were obtained by lentivirus transfection.The MTT cell proliferation experiment showed that the cell proliferation rate of the high expression group was faster(P=0.003),and the cell proliferation ability of the knockdown group was decreased(P=0.022);The apoptosis test of flow cytometry showed that the apoptosis of the high expression group decreased(P=0.003),and the apoptosis of the knockdown group increased(P=0.001);Transwell invasion and migration experiments showed that the invasion(P=0.01)and migration(P=0.006)of cells in the high expression group increased,while the invasion(P=0.005)and migration(P=0.001)of cells in the knockdown group decreased.Conclusion:GSDMD can be used as a potential biomarker and therapeutic target to evaluate the occurrence,development and prognosis of renal clear cell carcinoma.