miR-194-5p/UBA2在肾透明细胞癌进展中的作用及机制研究OA

Objective:To investigate the expression pattern of miR-194-5p in clear cell renal cell carcinoma(ccRCC)and its effects on malignant biological behaviors of tumor cells,and to validate the targeted regulatory relationship between miR-194-5p and UBA2.Methods:The expression of miR-194-5p in ccRCC tissues and cell lines was measured by real time RT-PCR.miR-194-5p agomir,miR-194-5p antagomir and corresponding negative control were transfected into 786-O and Caki-1 cells,and the expression of UBA2 protein and mRNA was detected.Cell proliferation,migration/invasion,and apoptosis were evaluated using CCK-8 assays,transwell assays,and flow cytometry,respectively.A dual-luciferase reporter assay was performed to verify the direct binding between miR-194-5p and the 3´UTR of UBA2.Results:miR-194-5p was significantly downregulated in ccRCC tissues and ccRCC cell lines.Up-regulation of miR-194-5p could reduce the expression of UBA2 protein and mRNA,inhibit cell proliferation,migration and invasion,and promote apoptosis.The dual-luciferase reporter assay showed that miR-194-5p specifically suppressed the reporter activity of constructs containing the wild-type UBA2 3´UTR,whereas no significant suppression was observed in UBA2 3´UTR 5´-UGUUACA-3´mutant constructs.Conclusion:Restoration of miR-194-5p expression can suppress the malignant biological behaviors of ccRCC cells by directly targeting and inhibiting UBA2.