| Objective: Renal cell carcinoma(RCC)is one of the most common malignancies in the urinary system,accounting for about 3% of the incidence of malignant tumors in adults.Renal clear cell carcinoma is the main phenotype of renal cell carcinoma,accounting for about 80%.Mi RNA is a single-stranded,endogenous non-coding small molecule RNA composed of about 22 nucleotides,which is relatively stable,and Mi RNA is involved in many biological processes.Although some mi RNAs have been found to be closely related to renal cell carcinoma,most of them are studies on single mi RNA markers.Since single mi RNA markers are often not highly specific,this study aims to look for multiple mi RNA combined markers to accurately predict the prognosis of patients with renal clear cell carcinoma.Methods: in this study,data sets related to renal clear cell carcinoma were downloaded from the cancer genome atlas(TCGA)database to analyze the differences in mi RNA expression between the two groups.We also obtained mi RNA expression profiles and clinical data sheets of 537 patients with renal clear cell carcinoma,and analyzed the survival of patients with renal clear cell carcinoma by Kaplan-Meier curve method,so as to screen out mi RNAs with potential biomarker characteristics.Target genes of differentially expressed mi RNAs were predicted by online mi RNA databases: mi RWalk,mi RDB,DIANA TOOL and mi RDIP.Then,the common target genes were used for gene ontology(GO)and gene enrichment analysis of Kyoto gene and genome encyclopedia(KEGG)to explore the biological processes and signaling pathways involved in or related to target genes.Differentially expressed mi RNAs were also verified by RT-q PCR in renal cell lines.Meanwhile,differential expression of mi RNA and clinicopathological features were also analyzed by chi-square analysis,COX univariate and multivariate analysis.Results: 1.In the TCGA data of renal clear cell carcinoma,72 adjust tissue samples and 539 renal clear cell carcinoma tissue samples were screened and included in the study for differential expression of mi RNA analysis.263 differentially expressed mi RNAs in renal clear cell carcinoma and adjacent tissues were screened,including 164 up-regulated mi RNAs and 99 down-regulated mi RNAs(P<0.05,|FC|>1).2.According to the expression profile chip and clinical data of renal clear cell carcinoma provided by the TCGA database,survival analysis was conducted on the differentially expressed mi RNA obtained in the previous step through the Kaplan-Meier curve,and it was found that patients with high expression of mi R-612,mi R-616-5p and mi R-760 had a short overall survival time,namely a poor prognosis.In the chi-square analysis,the expression of mi R-612 was correlated with histological grade.The expression of mi R-616-5p was correlated with T stage histological grade and pathological stage.The expression of mi R-760 was correlated with T stage,N stage,histological grade and pathological stage.3.Cox univariate and multivariate analyses showed that these three combined mi RNA markers have the potential to become important prognostic markers for renal clear cell carcinoma.In Cox univariate analysis,the patients were divided into high and low risk expression groups according to the risk score of the combined mi RNA,and the survival curve showed that it was related to the prognosis and survival time of patients.Cox univariate analysis showed that pathological stage(HR=3.784,P < 0.001),T stage(HR=2.972,P < 0.001),M stage(HR=4.186,P < 0.001),histological grade(HR=2.653,P < 0.001),and three mi RNA associated markers(HR=1.546,P=0.005)were significantly correlated with the overall survival of patients with renal clear cell carcinoma.Multivariate analysis showed that T stage(HR=1.794,P=0.002),M stage(HR=2.080,P=0.036),histological grade(HR=2.400,P < 0.001)and three mi RNA associated markers(HR=1.499,P=0.012)were independent predictors of prognosis in patients with renal clear cell carcinoma.4.The three mi RNAs had 241 common target genes,which were analyzed for gene function enrichment.Biological process and molecular function analysis showed that target genes were related to gene silencing,cell protein metabolism,hypoxia,nucleic acid binding,and the signal activity of transcription factor serine/threonine kinase receptor.Cell component analysis showed that target genes were involved in nucleosome,nucleoplasm,cytoplasmic matrix,chromatin,protein complex,transcription factor complex,exosome,cell-cell adhesion link,etc.KEGG pathway analysis showed that target genes were involved in p53,hypoxia-inducible factor 1,AMPK,renal cell carcinoma,and transcriptional disorders in tumors.5.Real-time fluorescence quantitative polymerase chain reaction(q RT-PCR)results showed that three mi RNAs(mi R-612,mi R-616-5p,mi R-760)were highly expressed in renal clear cell carcinoma cell lines.Conclusion: 1.Mi R-612,mi R-616-5p and mi R-760 were associated with the overall survival,TNM stage,pathological stage and other clinical characteristics of patients with renal clear cell carcinoma.Patients in the high-risk group combined with three mi RNAs had poor prognosis.2.The common target genes of three associated mi RNAs(mi R-612,mi R-616-5p,and mi R-760)are involved in the biological processes and signaling pathways related to the development of renal clear cell carcinoma.3.Three associated mi RNA markers(mi R-612,mi R-616-5p,mi R-760)can be used as potential markers for the development and prognosis of renal clear cell carcinoma. |
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