Identification of Prognostic Markers of DNA Damage and Oxidative Stress in Diagnosing Papillary Renal Cell Carcinoma Based on High-Throughput Bioinformatics Screening.

Abstract

Papillary renal cell carcinoma (pRCC) is the second most common histological subtype of adult kidney tumors, with a poor prognosis due to limited understanding of the disease mechanism. Herein, we have performed high-throughput bioinformatic screening to explore and identify potential biomarkers of DNA damage and oxidative stress for pRCC. RNA sequencing data related to pRCC were downloaded from the TCGA database, and differentially expressed genes (DEG) were identified by a wide variety of clustering and classification algorithms, including self-organized maps (SOM), artificial neural networks (ANN), support vector machines (SVM), fuzzy logic, and hyphenated techniques such as neuro-fuzzy networks. Then DAVID and STRING online biological information tools were used to analyze functional enrichment of the regulatory networks of DEG and construct a protein-protein interaction (PPI) network, and then the Cytoscape software was used to identify hub genes. The importance of key genes was assessed by the analysis of the Kaplan-Meier survival curves using the R software. Lastly, we have analyzed the expression of hub genes of DNA damage and oxidative stress (BDKRB1, NMUR2, PMCH, and SAA1) in pRCC tissues and adjacent normal tissues, as well as the relationship between the expression of hub genes in pRCC tissues and pathological characteristics and prognosis of pRCC patients. A total of 1,992 DEGs for pRCC were identified, with 1,142 upregulated ones and 850 downregulated ones. The DEGs were significantly enriched in activities including DNA damage and oxidative stress, chemical synaptic transmission, an integral component of the membrane, calcium ion binding, and neuroactive ligand-receptor interaction. cytoHubba in the Cytoscape software was used to determine the top 10 hub genes in the PPI network as BDKRB2, NMUR2, NMU, BDKRB1, LPAR5, KNG1, LPAR3, SAA1, MCHR1, PMCH, and NCAPH. Furthermore, the expression level of hub genes BDKRB1, NMUR2, PMCH, and SAA1 in pRCC tissues was significantly higher than that in the adjacent normal tissues. Meanwhile, the expression level of hub genes BDKRB1, NMUR2, PMCH, and SAA1 in pRCC tissues was significantly positively correlated with tumor stage, lymph node metastasis, and the histopathology grade of pRCC. In addition, high expression levels of hub genes BDKRB1, NMUR2, PMCH, and SAA1 were associated with a poor prognosis for patients with pRCC. Univariate and multivariate analyses showed that the expression of hub genes BDKRB1, NMUR2, PMCH, and SAA1 were independent risk factors for the prognosis of patients with pRCC. The results of this analysis suggested that BDKRB1, NMUR2, PMCH, and SAA1 might be potential prognostic biomarkers and novel therapeutic targets for pRCC.