MicroRNA-200a Regulates The Epithelial-Mesenchymal Transition Of Bladder Cancer Cells Through Targeting β-catenin

Objective: Through related in vitro biological behavior experiments,the role of miR-200 a in bladder cancer cell 5637 was explored.And explore the molecular mechanism of miR-200 a in the occurrence and development of bladder cancer.Methods: Transfection of mimetics(miR-200 a mimics,miR-200 a inhibitor,miR-NC)in bladder cancer cells 5637,and the transfection efficiency was evaluated by PCR.Tetramethylazoazole blue(MTT)method,Trace tests and Transwell cells were used to test the effects of miR-200 a on cell proliferation,metastasis and invasion of bladder cancer cells 5637.TOP / FOP flash luciferase method was used to identify the effect of miR-200 a on the activity of Wnt / β-catenin signal pathway.Luciferase method was used to determine the miR-200 a on the 3 ’untranslated region(3’-UTR).Western blot determined the effect of miR-200 a on downstream molecules of Wnt / β-catenin signal and molecules related to epithelial-mesenchymal transition(EMT).Results: Related experiments were performed using transfected cells.The results of the MTT method showed that the cell proliferation rate of the miR-200 a mimic group was significantly lower than those of the other groups;the scratch test results showed that the number of migrating cells in the mimic group was significantly less than that of the inhibitor group,negative control group,Irrelevant sequence group;the results of the Transwell chamber showed that the mimic group had a weaker cell invasion ability than the inhibitor group,negative control group and irrelevant sequence group.These results indicate that miR-200 a inhibits related biological functions of bladder cancer cell 5637.TOP / FOP flash luciferase method results showed that miR-200 a reduced the activity of the Wnt / β-catenin signaling pathway;luciferase method demonstrated that miR-200 a has a 3 ’untranslated region(3’-UTR)has a targeting effect;Western blot results show that miR-200 a overexpression down-regulates the levels of MMP9 and Cyclin D1 downstream molecules of Wnt / β-catenin signal,while miR-200 a inhibition increases their expression.These results indicate that miR-200 a can inhibit the biological function of bladder cancer cell 5637 by targeting the Wnt / β-catenin signaling pathway.Another part of the data from Western blot showed that miR-200 a overexpression induced downregulation of ZEB1,ZEB2 and N-cadherin,and up-regulation of E-cadherin,which indicated that miR-200 a played a role in the epithelial-mesenchymal transition(EMT)of bladder cancer.Conclusion: miR-200 a may play a role as a tumor suppressor gene in bladder cancer,and by targeting β-catenin,it inhibits biological functions such as proliferation,migration and invasion of bladder cancer cell 5637.And miR-200 a can play a role in the EMT of bladder cancer by regulating the genes related to the epithelial-mesenchymal transition(EMT)of bladder cancer.The above results indicate that miR-200 a is expected to become a new biomarker and therapeutic target for the diagnosis and treatment of bladder cancer.