High-risk neuroblastoma, as a lethal pediatric tumor, has unfavorable clinical outcomes, and accounts for ~15% of childhood cancer‐related mortality. However, the biomarkers for treatment prognosis have remained largely elusive. The present study sought to identify potential genes associated with the progression and prognosis of high-risk neuroblastoma.
The gene expression profiles of neuroblastoma were downloaded from the Gene Expression omnibus (GEO) database, and samples from GSE62564 and GSE16476 were applied as the training set and validation set, respectively. Bioinformatic analysis were performed by R. Subsequently, the biological functions of key gene were investigated by transwell, western blot, and flow cytometry analysis. miRNA that regulated the expression of key gene were predicted and the bind site were validated by luciferase reporter analysis and Q-PCR in children with high-risk neuroblastoma.
Analysis shown that 120 differentially expressed genes (DEGs) in the blue module were identified to be significantly related to high-risk neuroblastoma by weighted correlation network analysis (WGCNA). After MCODE and Cytohubba algorithm analyses, 26 genes were recognized as hub genes. Subsequently, univariate COX and LASSO regression analyses identified a six-gene signature (NCAPG, UBE2C, MELK, CCNA2, KIF15, and BIRC5) and constructed a prognostic model. The expression of these six genes in high-risk neuroblastoma children was significantly higher than that in non-high-risk neuroblastoma children. These signature genes and the prognostic model demonstrated strong prognostic capability using Kaplan-Meier (KM) curves. Subsequently, NCAPG was selected as key gene and the results indicated that knockdown NCAPG suppressed the migration and invasion, increased the apoptosis rate and decreased the Bcl-2 level in neuroblastoma cells. In addition, the luciferase reporter analysis method showed that miR-495-3p can negatively regulate the expression of NCAPG. Q-PCR indicated that miR-495-3p highly expressed in children with high-risk neuroblastoma.
Our findings identified that miR-495-3p/ NCAPG could serve as prognostic biomarkers of high-risk neuroblastoma and may advance the understanding of the molecular pathogenesis.