| Objective: Clear cell renal cell carcinoma(cc RCC)has an insidious clinical onset,about30% of patients are found incidentally,and about 25% of patients have metastasized when diagnosed.In addition,40% of patients who underwent radical nephrectomy relapsed and developed metastatic kidney cancer after surgery.Patients with advanced clear cell renal cell carcinoma have a high mortality rate,with a five-year survival rate of only 18%.Therefore,it is necessary to clarify the mechanism related to the occurrence and development of clear cell renal cell carcinoma for its early diagnosis and treatment.Inhibitor of growth family member 5(ING5)can regulate the cell cycle,apoptosis,epithelial-mesenchymal transition,and chromatin remodeling,which is involved in a variety of malignant tumors’ abnormal expression.The role of ING5 in clear cell renal cell carcinoma has not been reported,so this study aims to explore the expression of ING5 in clear cell renal cell carcinoma and its relationship with the clinicopathological characteristics of clear cell renal cell carcinoma through bioinformatics analysis and clinical tissue samples The relationship between them,and its influence on the malignant biological behavior of human renal clear cell carcinoma A498 cells was explored through cell experiments.Methods: 1.Using bioinformatics analysis,compare the expression of ING5 mRNA and protein in renal clear cell carcinoma tissues and adjacent tissues.2.Extracting the mRNA and protein of clear cell renal cell carcinoma tissue and adjacent tissue,and compare the expression of ING5 mRNA and protein in clear cell renal cell carcinoma tissue and adjacent tissue by RT-qPCR and Western blot.3.Using immunohistochemical staining techniques,compare the positive rate of ING5 protein expression in clear cell renal cell carcinoma tissues and adjacent tissues,and analyze the relationship between the expression of ING5 protein and the clinicopathological features of clear cell renal cell carcinoma patients.4.Using renal clear cell carcinoma A498 cells as tool cells,overexpressing ING5 in A498 cells,and using CCK-8 cell proliferation experiments,wound healing assay,and Transwell migration and invasion assay to clarify the effect of ING5 overexpression on the malignant biology of A498 cells behavioral impact.5.Using Western blot to explore the effect of ING5 overexpression on the expression of A498 cell proliferation and migrationrelated proteins.Results: 1.The results of bioinformatics analysis showed that the expression of ING5 mRNA and protein in renal clear cell carcinoma tissues was lower than that in adjacent tissues(P<0.05).2.The results of RT-qPCR and Western blot after extraction of mRNA and protein from clinical samples showed that the expression of ING5 mRNA and protein in clear cell renal cell carcinoma tissue was lower than that in adjacent tissue(P<0.05).3.The results of immunohistochemical staining showed that the positive rate of ING5 protein expression in clear cell renal cell carcinoma was lower than that in adjacent tissues;the correlation analysis results of ING5 protein expression combined with the patient’s clinical data showed that the expression of ING5 protein was related to the patient’s age(P=0.016)It was positively correlated and negatively correlated with the patient’s BMI index(P=0.016).4.The results of the CCK-8 proliferation experiment showed that the proliferation rate of ING5 overexpressed A498 cells was higher than that of control cells.From the 16 th hour,there was a difference in the proliferation rate between the two(P<0.05);the results of the wound healing assay showed that ING5 overexpressed A498 The wound healing speed of the cells was significantly increased compared with the control cells(P<0.05);in the Transwell migration and invasion assay,the number of ING5 overexpressed A498 cells passing through the chamber was significantly higher than that of the control cells(P<0.05).5.The results of Western blot showed that the protein expression of p-AKT,p-GSK-3β,SNAI1,and Cyclin D1 in ING5 overexpressed A498 cells increased compared with control cells(P<0.05),while the expression of E-Cadherin protein decreased compared with control cells(P<0.05).Conclusion: 1.ING5 mRNA and protein expression decreased in clear cell renal cell carcinoma.ING5 protein expression was positively correlated with age in patients with clear cell renal cell carcinoma,and negatively correlated with body mass index.2.Overexpression of ING5 can promote the proliferation,migration,and invasion of cc RCC A498 cells.3.Overexpression of ING5 down-regulates E-Cadherin to promote EMT in A498 cells. |
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