Identification of SSBP1 as a ferroptosis-related biomarker of glioblastoma based on a novel mitochondria-related gene risk model and in vitro experiments

J Transl Med. 2022 Sep 30;20(1):440. doi: 10.1186/s12967-022-03657-4.

Abstract

Background: Glioblastoma (GBM) is the most common primary malignant brain tumor that leads to lethality. Several studies have demonstrated that mitochondria play an important role in GBM and that mitochondria-related genes (MRGs) are potential therapeutic targets. However, the role of MRGs in GBM remains unclear.

Methods: Differential expression and univariate Cox regression analyses were combined to screen for prognostic differentially-expressed (DE)-MRGs in GBM. Based on LASSO Cox analysis, 12 DE-MRGs were selected to construct a risk score model. Survival, time dependent ROC, and stratified analyses were performed to evaluate the performance of this risk model. Mutation and functional enrichment analyses were performed to determine the potential mechanism of the risk score. Immune cell infiltration analysis was used to determine the association between the risk score and immune cell infiltration levels. CCK-8 and transwell assays were performed to evaluate cell proliferation and migration, respectively. Mitochondrial reactive oxygen species (ROS) levels and morphology were measured using a confocal laser scanning microscope. Genes and proteins expression levels were investigated by quantitative PCR and western blotting, respectively.

Results: We identified 21 prognostic DE-MRGs, of which 12 DE-MRGs were selected to construct a prognostic risk score model for GBM. This model presented excellent performance in predicting the prognosis of patients with GBM and acted as an independent predictive factor. Functional enrichment analysis revealed that the risk score was enriched in the inflammatory response, extracellular matrix, and pro-cancer-related and immune related pathways. Additionally, the risk score was significantly associated with gene mutations and immune cell infiltration in GBM. Single-stranded DNA-binding protein 1 (SSBP1) was considerably upregulated in GBM and associated with poor prognosis. Furthermore, SSBP1 knockdown inhibited GBM cell progression and migration. Mechanistically, SSBP1 knockdown resulted in mitochondrial dysfunction and increased ROS levels, which, in turn, increased temozolomide (TMZ) sensitivity in GBM cells by enhancing ferroptosis.

Conclusion: Our 12 DE-MRGs-based prognostic model can predict the GBM patients prognosis and 12 MRGs are potential targets for the treatment of GBM. SSBP1 was significantly upregulated in GBM and protected U87 cells from TMZ-induced ferroptosis, which could serve as a prognostic and therapeutic target/biomarker for GBM.

Keywords: Ferroptosis; Glioblastoma; Mitochondria; SSBP1; Temozolomide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers
  • DNA-Binding Proteins / metabolism
  • Ferroptosis*
  • Gene Expression Regulation, Neoplastic
  • Glioblastoma* / pathology
  • Humans
  • Mitochondria / metabolism
  • Mitochondrial Proteins / genetics
  • Reactive Oxygen Species / metabolism
  • Sincalide / genetics
  • Sincalide / metabolism
  • Sincalide / therapeutic use
  • Temozolomide / pharmacology

Substances

  • Biomarkers
  • DNA-Binding Proteins
  • Mitochondrial Proteins
  • Reactive Oxygen Species
  • SSBP1 protein, human
  • Sincalide
  • Temozolomide