Screening And Identification Of MiRNA Targeting And Regulating Bladder Cancer Metastasis Suppressor Gene RhoGDI2

OBJECTIVE: Screen and identify miRNAs targeting the regulation ofRhoGDI2gene in bladder cancer.METHODS: First,download the gene expression and miRNA sequencing data and methylation data of bladder cancer through The Cancer Genome Atlas(TCGA).The online website predicts that miRNA withRhoGDI2has binding sites.Then use the bladder cancer data of TCGA database to analyze the correlation of miRNA-m RNA and methylation-m RNA respectively to find out the possible regulation mode of RhoGDI2;construct a plasmid vector withRhoGDI2sequence and co-transfect it with pre-selected miRNA mimics Double luciferase experiments were carried out in 293 T cells to detect their molecular binding ability;Western blot was used to detect the protein expression ofRhoGDI2and RAC1 in 4 different aggressive bladder cancer cells,and to compare the bladder cancer under differentRhoGDI2expression Cell biological behavior.RESULTS: Through preliminary bioinformatics analysis,we believe that post-transcriptional regulation of miRNA is more likely to be the potential regulation mode of RhoGDI2,and preliminary screening of hsa-miR-34c-5p and hsa-miR-375 as pre-selected miRNA;in dual fluorescence In the enzyme experiment,the 3’UTR plasmid ofRhoGDI2and hsa-miR-375 mimic were cotransfected and the fluorescence activity of firefly was significantly decreased compared with other groups(P<0.05),and there was a binding site between the two.No binding site was detected between hsa-miR-34c-5p and RhoGDI2;we found that in 4 types of bladder cancer cells,there was a significant positive correlation betweenRhoGDI2and RAC1 protein expression;RT4 and SW780RhoGDI2expression was significantly higher T24 and UMUC3.At the same time,RT4 and SW780 are significantly weaker than T24 and UMUC3 in cell proliferation and invasion ability.CONCLUSION: Patients with higher RhoGDI2 expression will have a better prognosis.Compared with the regulation of methylation,the decreased expression ofRhoGDI2in bladder cancer is more likely to be caused by the posttranscriptional regulation of miRNA.The dual luciferase experiment confirmed that hsa-miR-375 andRhoGDI2have the molecular binding ability.RhoGDI2 and RAC1 have a strong positive correlation,and bladder cancer cells with high expression ofRhoGDI2have significantly lower cell proliferation and invasion ability.