Hirudin suppresses ovarian cancer cell proliferation and glycolysis through inhibiting the NF-κB/HK3 axis

Background: Ovarian Cancer (OC) is a highly lethal malignancy, and chemoresistance remains a major obstacle to effective treatment. This study aims to investigate the therapeutic mechanisms of hirudin in OC, with the goal of identifying novel strategies to overcome chemoresistance.

Methods: The CCK8 assay was used to assess the viability of OC cells treatment with various concentrations of hirudin. Colony formation assays were conducted to evaluate cell proliferation, while wound healing and Transwell assays were used to examine cell migration and invasion, respectively. To investigate the potential molecular mechanisms underlying hirudin's effects on OC, network pharmacology analysis was performed by integrating data from SuperPred, TargetNet, GeneCards, OMIM, and DisGeNET to identify candidate target genes associated with both hirudin and OC. Importantly, dual-luciferase reporter assays were carried out to further evaluate the regulatory effects of hirudin on the transcriptional activity of key target genes.

Results: We found that hirudin inhibits the proliferation, migration, and invasion of OC cells in a concentration-dependent manner. Network pharmacology analysis identified nuclear factor-κB (NF-κB) as a potential key molecular target of hirudin. Mechanistically, hirudin suppresses glycolytic activity by inhibiting NF-κB to downregulate Hexokinase 3 (HK3) expression in OC cells. Consequently, both the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were significantly reduced.

Conclusion: This study provides a theoretical foundation for elucidating the molecular mechanisms underlying the anti-tumor effects of hirudin in OC and suggests its potential as a candidate for metabolism-targeted therapy.