Temozolomide and ruxolitinib combination modulates miRNA-associated regulatory networks in glioblastoma stem cells

Glioblastoma is an aggressive primary brain tumor characterized by high recurrence rates and resistance to standard therapies, including temozolomide (TMZ). Emerging evidence suggests that microRNAs (miRNAs) play a critical role in regulating treatment response and tumor progression. In this study, we investigated the effects of TMZ and the JAK inhibitor ruxolitinib, alone and in combination, on miRNA expression profiles in glioblastoma cancer stem cells. miRNA expression levels were analyzed using quantitative real-time PCR, and differential expression was evaluated using the 2^-ΔΔCt method. TMZ treatment increased the expression of oncogenic miRNAs, including hsa-miR-19a-3p, hsa-miR-221-3p, and hsa-miR-10a-5p, whereas ruxolitinib treatment reduced their expression levels. Combination treatment attenuated the TMZ-induced upregulation of these miRNAs. Bioinformatics analyses were performed to identify target genes and associated signaling pathways. Predicted targets were obtained using TargetScan and further supported by experimentally validated interactions from miRTarBase. KEGG pathway enrichment analysis revealed significant associations with cancer-related pathways, including PI3K-Akt, MAPK, Wnt, and Hippo signaling pathways. Network analysis highlighted key genes such as BCL2L11, PTEN, and SOCS family members. Overall, our findings suggest that TMZ may induce oncogenic miRNA expression as part of an adaptive response, while ruxolitinib may counteract this effect. Targeting miRNA-mediated regulatory networks in combination with pathway inhibition may represent a promising strategy to overcome therapeutic resistance in glioblastoma.