E2F1-Driven CENPM Expression Promotes Glycolytic Reprogramming and Tumorigenicity in Glioblastoma

Background: While historically linked to chromosome segregation, centromere proteins (CENPs) are increasingly implicated in cancer biology. Although other CENPs have been studied in various cancers, including Glioblastoma (GBM), the less-explored CENPM demonstrates intriguing potential to influence tumor cell proliferation, invasion, and metabolic reprogramming - crucial factors in GBM aggressiveness. Methods: Initial insights into CENPM expression patterns in GBM were obtained through GEPIA database analysis (TCGA/GTEx datasets). Our in vitro model incorporated LN229 and U251 GBM cell lines. We modulated CENPM expression through shRNA-mediated knockdown and overexpression (pcDNA3.1 vectors), using Lipofectamine 2000 delivery. RT-qPCR and Western blotting assays quantified CENPM levels at mRNA and protein levels. GBM cell viability and proliferation were assessed via CCK-8 and EdU assays, respectively, while Transwell assays evaluated invasion. Changes in cellular metabolism were analyzed by measuring glucose consumption, lactate production, and ATP levels. To evaluate E2F1's regulatory impact on CENPM, we used luciferase reporter assays. Finally, an in vivo BALB/c nude mouse xenograft model helped us assess the effect of CENPM modulation on tumor growth. Results: Our analysis demonstrated significant CENPM upregulation in GBM, confirmed by GEPIA and RT-qPCR, suggesting its potential as a therapeutic target. CENPM knockdown dramatically reduced GBM cell proliferation and invasion, accompanied by altered EMT marker expression. This highlights CENPM's central role in GBM progression. Conversely, CENPM overexpression promoted growth and EMT, further implicating it in aggressive tumor behavior. We observed CENPM's influence on glycolysis, with reduced activity following knockdown and increased activity with overexpression. Importantly, the glycolytic inhibitor 2-DG reversed these effects, hinting at glycolytic dependency in CENPM-modulated cells. We also established that E2F1 interacts with the CENPM promoter, representing a likely regulatory mechanism. Finally, in vivo studies confirmed that CENPM knockdown significantly hinders tumor growth, strongly supporting its critical role in GBM. Conclusion: Our findings highlight a novel role for CENPM in promoting GBM progression through metabolic reprogramming and enhancing tumor cell invasion. The E2F1-CENPM axis emerges as a critical mediator of these processes, suggesting that targeting this interaction could offer a new therapeutic approach for GBM treatment.