基于TCGA数据库分析DTX2在肾透明细胞癌组织中表达的临床意义及其对肾癌细胞增殖、迁移与侵袭的影响OA北大核心CSTPCD

Objective:To analyze the expression level and clinical significance of Deltex E3 ubiquitin ligase 2(DTX2)in clear cell renal cell carcinoma(ccRCC)based on TCGA database,and explore the effect of DTX2 on the proliferation,migration and invasion of ccRCC cells.Methods:TIMER database was utilized to analyze the expression level of DTX2 in pan-cancer tissues,while UALCAN database was used for further verification of the differences in mRNA and protein expressions of DTX2 in ccRCC and adjacent tissues.TCGA-ccRCC cohort dataset in UALCAN database was employed to examine the correlation between DTX2 expression in ccRCC and clinicopathological features of patients.The correlation between DTX2 expression and the prognosis of ccRCC patients was analyzed using K-M plot database.Using DAVID database,gene ontology(GO)analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed to explore DTX2-related genes.The expression levels of DTX2 gene in human embryonic kidney 293(HEK293)cells and ccRCC A498 and Caki-1 cells were detected by qPCR.SiRNA technology was employed to transfect DTX2 siRNA and its negative control plasmids into ccRCC A498 and Caki-1 cells.CCK-8 cell proliferation assay,plate clone formation assay,scratch wound assay,and Transwell migration assay were performed to detect respectively the effects of knockdown DTX2 on the proliferation,migration and invasion of ccRCC cells.Results:Analysis of TCGA database showed that,compared with adjacent tissues,both DTX2 mRNA and protein were highly expressed in ccRCC tissues(all P<0.01).The expression level of DTX2 was associated with the pathological stage,clinical grade,different subtype,and lymph node metastasis of ccRCC patients(all P<0.01).High DTX2 expression was correlated with poor prognosis in patients(all P<0.01).The results of GO function analysis and KEGG pathway enrichment analysis and showed that genes related to DTX2 expression were mainly involved in biological processes such as proteasome-mediated ubiquitin-dependent protein catabolic process,and these genes were primarily enriched in tumor-related signaling pathways such as the mTOR signaling pathway(all P<0.05).The results of in vitro cell experiments showed that the expression levels of DTX2 in A498 and Caki-1 cells were higher than those in HEK293 cells;knockdown of DTX2 expression significantly lowered the proliferation,migration and invasion abilities of A498 and Caki-1 cells(all P<0.01).Conclusion:High expression of DTX2 is observed in ccRCC tissues and cells,and its high expression is associated with poor prognosis in patients.Knockdown of DTX2 expression may inhibit the proliferation,migration and invasion of ccRCC cells.DTX2 is expected to become a new biomarker and therapeutic target for ccRCC.