Molecular Mechanism Of MiR-489-3p/Twist Inhibiting EMT In Clear Cell Renal Cell Carcinoma

Objective:The aim of this experiment is to investigate the expression levels of mi R-489-3p and Twist in renal cancer tissues and cells,and to explore the molecular mechanism of mi R-489-3p Twist pathway regulating EMT in renal clear cell carcinoma,providing a theoretical basis for the treatment and prognosis of renal clear cell carcinoma.Methods:A total of 20 samples of renal clear cell carcinoma tissues and adjacent tissues confirmed by pathology after radical nephrectomy and surgery were collected from the People’s Hospital of Guizhou Province.The expression level of mi R-489-3p was detected using real-time fluorescence quantitative PCR(Quantitative RT-PCR)method;Cultivate one normal renal tubular epithelial cell line(HK-2)and two human transparent renal cell carcinoma(Clear cell renal cell carcinoma,cc RCC)cell lines(OSRC2,786-O)and detect the expression of mi R-489-3p at the cellular level;Select the 786-O renal clear cell carcinoma cell line for q PCR and Western blotting methods to detect the expression levels of mi R-489-3p on Twist and EMT related proteins;Cell clone assay,scratch assay,and Transwell assay were used to detect cell proliferation,migration,and invasion abilities;The targeted regulation of mi R-489-3p on Twist was determined by the Dual Luciferase Reporter Assay.Results:The q PCR experiment results showed that the expression level of mi R-489-3p in renal clear cell carcinoma tissues was lower than that in normal adjacent tissues,with statistical significance(P<0.05);The expression level of mi R-489-3p was low in renal cancer cell lines 786-O and OSRC2,and the difference was statistically significant compared to the normal renal tubular epithelial cell line HK-2(P<0.05).After successful transfection of mi R-489-3p mimics and mi R-489-3p inhibitor into the786-O cell line,cell cloning,scratch experiments,and Transwell experiments were performed.The results showed that compared with the NC mimics group,the proliferation,invasion,and migration abilities of 786-O cells in the mi R-489-3p mimics group were significantly reduced;The proliferation,invasion,and migration abilities of 786-O cells in the mi R-489-3p inhibitor group were significantly enhanced.The results of q PCR showed that Twist was highly expressed in 786-O cells,and the difference was statistically significant compared to HK-2 cells(P<0.05);The results of double Luciferase reporter gene experiment showed that mi R-489-3p combined with Twist 3’UTR,and there was a targeted regulation relationship.Conclusions:The expression level of mi R-489-3p is low in tissues and cells of clear cell renal cell carcinoma patients;mi R-489-3p can inhibit the proliferation,invasion,and migration of cc RCC cells;mi R-489-3p may be targeted to regulate twist,thereby inhibiting the occurrence and development of renal cell carcinoma.