Gene expression profiling and the isocitrate dehydrogenase mutational landscape of temozolomide‑resistant glioblastoma

Abstract

Glioblastoma multiforme (GBM) is an aggressive brain cancer that occurs more frequently than other brain tumors. The present study aimed to reveal a novel mechanism of temozolomide resistance in GBM using bioinformatics and wet lab analyses, including meta‑Z analysis, Kaplan‑Meier survival analysis, protein‑protein interaction (PPI) network establishment, cluster analysis of co‑expressed gene networks, and hierarchical clustering of upregulated and downregulated genes. Next‑generation sequencing and quantitative PCR analyses revealed downregulated [tyrosine kinase with immu‑ noglobulin and epidermal growth factor homology domains 1 (TIE1), calcium voltage‑gated channel auxiliary subunit α2Δ1 (CACNA2D1), calpain 6 (CAPN6) and a disintegrin and metal‑ loproteinase with thrombospondin motifs 6 (ADAMTS6)] and upregulated [serum amyloid (SA)A1, SAA2, growth differen‑ tiation factor 15 (GDF15) and ubiquitin specific peptidase 26 (USP26)] genes. Different statistical models were developed for these genes using the Z‑score for P‑value conversion, and Kaplan‑Meier plots were constructed using several patient cohorts with brain tumors. The highest number of nodes was observed in the PPI network was for ADAMTS6 and TIE1. The PPI network model for all genes contained 35 nodes and 241 edges. Immunohistochemical staining was performed using isocitrate dehydrogenase (IDH)‑wild‑type or IDH‑mutant GBM samples from patients and a significant upregulation of TIE1 (P<0.001) and CAPN6 (P<0.05) protein expres‑ sion was demonstrated in IDH‑mutant GBM in comparison with IDH‑wild‑type GBM. Structural analysis revealed an IDH‑mutant model demonstrating the mutant residues (R132, R140 and R172). The findings of the present study will help the future development of novel biomarkers and therapeutics for brain tumors.