Collagen type I alpha 1 induces radio-resistance in hypoxic glioblastoma cells by promoting autophagy.

AIM: To investigate how collagen type I alpha 1 gene (COL1A1) affects hypoxic glioblastoma (GBM) cell radiosensitivity and explore underlying mechanisms.

MAIN METHODS: We used GBM patients' bulk and single cell RNA sequencing (RNA-seq and scRNA-seq) data along with clinical data from various public databases to assess COL1A1 expression and its cell subgroups, with the Kaplan-Meier method for survival analysis. We used bioinformatics to identify correlations between COL1A1 and hypoxia-related pathways and fluorescence-based real-time quantitative polymerase chain reaction and immunoblotting to detect COL1A1 expression changes in GBM cells. Stable GBM cells with COL1A1 overexpression or knockdown were established using lentiviral vectors. We used CCK-8 assay, flow cytometry, and plate colony formation assay to evaluate GBM cells' proliferation, apoptosis, and radiosensitivity, respectively. We assessed autophagic flux with mRFP-GFP-LC3 dual fluorescence labeling and evaluated expression of apoptosis-related proteins (PARP, LC3B, P62) and DNA repair protein γ-H2Ax through immunoblotting.

KEY FINDINGS: In GBM patients, COL1A1 demonstrated universally strong expression which was significantly associated with shortened survival. In scRNA-seq data, COL1A1 is associated with a distinct cell subgroup whose functions are enriched in epithelial-mesenchymal transition (EMT), angiogenesis, protein secretion, and DNA repair. In GBM cells, irradiation and hypoxia increased COL1A1 expression; under hypoxic conditions, COL1A1 overexpression promoted proliferation. However, in hypoxic M059K cells, COL1A1 knockdown strongly inhibited growth and autophagy but increased radiosensitivity and apoptosis. Conversely, COL1A1 overexpression increased radioresistance and autophagy but inhibited apoptosis in hypoxic U251MG cells.Blocking the autophagy pathway weakened the regulation of radiosensitivity by COL1A1 in hypoxic GBM cells.