Background Lung cancer is the most common cancer worldwide and remains the leading cause of cancer-related mortality.A method that can be used for the early detection and diagnosis of non-small-cell lung cancer (NSCLC) is urgently needed.Materials and Methods We identified differentially expressed genes based on RNA sequencing data for NSCLC from The Cancer Genome Atlas (TCGA) database.Then,we conducted receiver operating characteristic (ROC) curve and survival analyses to determine which genes had diagnostic and/or prognostic value.Results A total of 24991 genes were included in the TCGA database,and a differential gene expression analysis was conducted for 971 NSCLC and 108 normal samples.There were 1709 differentially expressed genes identified using three methods.It was noted that 29 of the 1709 genes were enriched in the Transcriptional misregulation in cancer pathway,such as HOXA11,HOXA10,HMGA2,MMP3,and MMP9.Based on the ROC curve and survival analyses,27 of the 1709 differentially expressed genes were with exact diagnostic and prognostic value for NSCLC,such as SLC2A1,LGR4,EMP2,CLDN5,PRC1,ANLN,GAPDH.GAPDH took the center position in the protein-protein interaction (PPI)network.Conclusion Twenty-seven differentially expressed genes with diagnostic and prognostic value were identified that can serve as potential biomarkers for NSCLC.