Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults, with a poor prognosis despite advances in treatment. This review summarizes recent progress in GBM pathology, molecular mechanisms, and biomarkers. GBM is characterized by high heterogeneity, involving complex genetic, epigenetic, and transcriptomic alterations that affect major signaling pathways. Molecular features of GBM are crucial for diagnosis, prognosis, and targeted therapy. The 2016 WHO classification of CNS tumors incorporated molecular markers into GBM classification, emphasizing the importance of molecular biomarkers. Recent advances include single-cell technologies, deep learning-based multi-omics data exploration, and novel 3D preclinical models, which have enhanced the molecular characterization of GBM. Key molecular features include TERT promoter mutations, PTEN deletion, EGFR amplification, and IDH mutation status. Epigenetic changes, such as MGMT promoter methylation, are important for prognosis and treatment response. Transcriptomic studies have identified distinct subtypes of GBM, such as Proneural, Proliferative, Mesenchymal, and Neural, which have different prognostic implications. Key signaling pathways involved in GBM include the RTK, TP53, and RB pathways. Clinically relevant biomarkers include IDH1, MGMT, and EGFR, which are important for prognosis and therapy. Challenges in GBM treatment include intratumoral heterogeneity and the blood-brain barrier, which hinder drug delivery. Future directions include the development of combination therapies, improved biomarker identification, and the application of precision medicine. This review highlights the importance of molecular characterization in GBM diagnosis, prognosis, and treatment.Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults, with a poor prognosis despite advances in treatment. This review summarizes recent progress in GBM pathology, molecular mechanisms, and biomarkers. GBM is characterized by high heterogeneity, involving complex genetic, epigenetic, and transcriptomic alterations that affect major signaling pathways. Molecular features of GBM are crucial for diagnosis, prognosis, and targeted therapy. The 2016 WHO classification of CNS tumors incorporated molecular markers into GBM classification, emphasizing the importance of molecular biomarkers. Recent advances include single-cell technologies, deep learning-based multi-omics data exploration, and novel 3D preclinical models, which have enhanced the molecular characterization of GBM. Key molecular features include TERT promoter mutations, PTEN deletion, EGFR amplification, and IDH mutation status. Epigenetic changes, such as MGMT promoter methylation, are important for prognosis and treatment response. Transcriptomic studies have identified distinct subtypes of GBM, such as Proneural, Proliferative, Mesenchymal, and Neural, which have different prognostic implications. Key signaling pathways involved in GBM include the RTK, TP53, and RB pathways. Clinically relevant biomarkers include IDH1, MGMT, and EGFR, which are important for prognosis and therapy. Challenges in GBM treatment include intratumoral heterogeneity and the blood-brain barrier, which hinder drug delivery. Future directions include the development of combination therapies, improved biomarker identification, and the application of precision medicine. This review highlights the importance of molecular characterization in GBM diagnosis, prognosis, and treatment.