Selection of reference genes and proteins for expression studies in pediatric gliomas.

A critical aspect of gene and protein expression analysis is the selection of stable reference genes and proteins. These references are essential for normalizing data and enabling accurate comparisons between samples. In today’s context, molecular typing is essential to enhance diagnostic precision and inform therapeutic decisions, further emphasizing the need for reliable reference molecules. In this study, we analyzed the expression levels of six reference gene candidates and seven proteins in pediatric glioma samples. Gliomas are the most common solid tumors found in the central nervous system of children and are typically classified into low-grade gliomas, optic pathway gliomas, and high-grade gliomas. We evaluated the expression levels of the six target genes using RefFinder web-based software. Our findings indicate that YWHAZ and GAPDH are the most reliable pairs of reference genes, while β-Actin and 14-3-3ζ serve as the most dependable reference proteins. Interestingly, the expression of putative housekeeping proteins in low-grade gliomas closely resembles that in high-grade gliomas but differs from the expression observed in optic pathway gliomas. Furthermore, we found no correlation between RNA amounts and protein levels, suggesting variations in how protein expression is regulated. Overall, the results show that the expression of reference genes in pediatric gliomas differs from what has been reported for adult gliomas. This research provides valuable insights into the molecular characteristics of pediatric gliomas and may have important implications for diagnosis and treatment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12672-025-04119-1.