Screening The Differentially Expressed Gene UBE2C In Breast Cancer Based On Bioinformatics And Its Functional Verification

Objective: To screen the differentially expressed genes in breast cancer,analyze their effects on breast cancer cell proliferation and invasion,and explore the molecular mechanisms that affect breast cancer cell proliferation.Methods: Breast cancer-related datasets were screened according to filter criteria in the Gene Expression Omnibus(GEO)database and The Cancer Genome Atlas(TCGA)database.Then differentially expressed genes(DEGs)were identified by Venn diagram analysis.Using these genes,we conducted the following analyses including Gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG),protein-protein interaction(PPI)and survival analysis and then validated the function of the hub gene UBE2 C by RT-q PCR,CCK-8,transwell and Western blot assays.Results: In total,151 DEGs were identified from the GEO and TCGA databases.GO analysis results demonstrated that the DEGs were significantly enriched for mitotic cell cycle process,lipid droplet and protein dimerization.KEGG analysis showed that PPAR signaling pathway,regulation of lipolysis in adipocytes and proximal tubule bicarbonate reclamation were the most significantly enriched in the signal transduction pathway category.The top three hub genes resulted from the PPI network were FOXM1,UBE2 C,and CDKN3.Survival analysis results showed close relationship between UBE2 C and breast cancer.CCK-8 and transwell assay results suggested that the proliferation and invasion of UBE2 C knockdown cells were significantly inhibited.Western blot results showed that the level of phosphorylated PTEN(p-PTEN)was obviously increased,while the levels of phosphorylated AKT(p-AKT)?phosphorylated mTOR(p-mTOR)and HIF-1? were dramatically reduced in the UBE2 C knockdown cell.Conclusion: UBE2 C can promote breast cancer proliferation by activating the AKT/mTOR signaling pathway.