Abstract
OBJECTIVES: To investigate the role of heat shock protein family B (small) member 1 (HSPB1) in regulating ferroptosis in glioma and to explore the underlying molecular mechanisms. METHODS: HSPB1 expression was analyzed in glioma cell lines. U251 glioma cells were transfected with HSPB1-targeting short hairpin RNA (shRNA). Cell proliferation, invasion, ferroptosis markers (iron accumulation, oxidative stress), and expression of BCL2-associated athanogene 3 (BAG3) were assessed using molecular and biochemical assays. BAG3 was further silenced or overexpressed to evaluate its interaction with HSPB1 in regulating ferroptosis. RESULTS: HSPB1 was markedly overexpressed in glioma cell lines. HSPB1 knockdown significantly suppressed U251 cell proliferation and invasion, while promoting ferroptosis via increased intracellular Fe(2+) levels and lipid peroxidation. BAG3 was identified as a downstream target of HSPB1. Silencing BAG3 replicated the anti-tumor and pro-ferroptotic effects of HSPB1 knockdown. Moreover, BAG3 overexpression partially rescued the effects of HSPB1 silencing, confirming its role in the HSPB1-mediated ferroptosis pathway. CONCLUSIONS: HSPB1 inhibits ferroptosis and promotes glioma cell survival, at least in part through BAG3 upregulation. Targeting the HSPB1-BAG3 axis may represent a novel therapeutic strategy and prognostic approach in glioma treatment.