The role of ALKBH5 in regulating proliferation and invasion in U251 glioblastoma cells.

INTRODUCTION: This study aimed to investigate the role of ALKBH5 in the proliferation, invasion, and MUC1 expression of U251 glioblastoma cells, using both in vitro and in vivo methodologies. MATERIAL AND METHODS: This study investigated the role of ALKBH5, an RNA demethylase, in regulating the proliferation, invasion, and ferroptotic sensitivity of U251 glioblastoma cells, with a focus on its interplay with MUC1, a transmembrane glycoprotein implicated in tumour progression and stress responses. RESULTS: Using in vitro and in vivo models, we demonstrate that ALKBH5 knockdown significantly inhibits U251 cell proliferation, invasion, and tumour growth, while its overexpression enhances these oncogenic properties. ALKBH5 regulates MUC1 expression, with MUC1 overexpression that rescues the inhibitory effects of ALKBH5 knockdown on cell viability and invasion. We uncover a novel link between ALKBH5 and ferroptosis, i.e. that ALKBH5 knockdown sensitises U251 cells to ferroptotic cell death through modulation of MUC1-mediated lipid peroxidation and iron metabolism pathways. In vivo studies reveal reduced tumour growth and Ki-67 expression in ALKBH5 knockdown models with decreased MUC1 levels in tumour tissues. CONCLUSIONS: The results highlight ALKBH5 as a critical regulator of glioblastoma progression and ferroptotic sensitivity, with MUC1 acting as a key downstream effector. Our findings provide new insights into the molecular mechanisms that drive glioblastoma aggressiveness and propose ALKBH5 and MUC1 as promising therapeutic targets for combating this lethal disease.