| Objective:The study was designed to explore the effects of hsa-miR-143 to the biological behaviours of renal clear cell carcinoma cell line 786-O.Besides,we aim to find and identify the target gene of hsa-miR-143.Furthermore/we intend to study the influence of hsa-miR-143 in the epithelial mesenchymal transition of renal clear cell carcinoma.Thus,we will realize the molecular mechanism of hsa-miR-143 in the carcinogenesis and development of renal clear cell carcinoma.Methods:1. Renal clear cell carcinoma cell line 786-0 transfected with hsa-miR-143 was evaluated for adhesion assay; 2. Transwell cell migration assay was performed to evaluate the ability of cell migration of 786-0 after transfecting with hsa-miR-143; 3. Bio-informatics software Targetscan was used for the prediction of target gene of hsa-miR-143 and luciferase assay was carried out for confirmation of the target gene.4. Western blot assay was used to verify the effect of hsa-miR-143 to the target gene and the epithelial mesenchymal transition proteins.Results:1.In vitro study,the number of adhering cells was significantly lower after transfecting with hsa-miR-143 than the NC group.2. The number of migrating cells also decreased after transfecting with hsa-miR-143,the difference is statistically significant.3.KRAS was validated to be a direct target gene of hsa-miR-143 by luciferase assay.4.After transfecting with hsa-miR-143,the expression of E-cadherin raised, the expression of KRAS,Vimentin and N-cadherin decrased.Conclusion:Hsa-miR-143 is a negative modulator for cell adhesion and migration in renal clear cell carcinoma. Hsa-miR-143 also inhibits the epithelial mesenchymal transition of renal clear cell carcinoma. Thus, hsa-miR-143 plays the role of a tumor suppressor gene in renal clear cell carcinoma.KRAS is a direct target gene of hsa-miR-143. Hsa-miR-143 is expected to be a new tumor marker and therapeutic target for renal clear cell carcinoma. |
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