Reynosin targets the PRDX6-AKT-WEE1 axis to suppress osteosarcoma and preserve the bone microenvironment.

BACKGROUND

Osteosarcoma (OS) is a formidable bone malignancy frequently associated with aggressive metastasis and limited therapeutic options. In this study, we investigated the anti-neoplastic potential of reynosin, a bioactive sesquiterpene lactone derived from Aucklandia lappa Decne., focusing on its ability to disrupt the Peroxiredoxin 6 (PRDX6)-mediated oncogenic network.

METHODS

Clinical specimen analysis was conducted via immunohistochemistry to evaluate PRDX6 and WEE1 expression in OS patient tissues. In vitro functional assays were performed on MG63 cells to assess proliferation, migration, invasion, and apoptosis. The GeneMANIA algorithm was employed to predict functional protein networks. Mechanistic validation was achieved through gain-of-function experiments utilizing stable overexpression of PRDX6. Furthermore, an ex vivo mouse calvarial bone model was utilized to simulate the tumor-bone microenvironment and evaluate bone formation.

RESULTS

Clinical analysis revealed a significant upregulation of PRDX6 and its downstream effector, WEE1, in OS patient tissues. In vitro assays demonstrated that reynosin treatment significantly hindered MG63 cell proliferation, migration, and invasion by triggering ROS-dependent mitochondrial apoptosis. GeneMANIA predicted a robust functional coupling within the PRDX6-AKT-WEE1 axis. Mechanistic investigations showed that reynosin-induced PRDX6 suppression led to the dephosphorylation of AKT and the subsequent downregulation of WEE1. Crucially, stable overexpression of PRDX6 effectively neutralized the pro-oxidative and anti-proliferative effects of reynosin by restoring p-AKT and WEE1 levels. In the ex vivo model, PRDX6-overexpressing MG63 cells significantly suppressed calvarial bone formation. Notably, reynosin treatment effectively reversed these deleterious effects, restoring bone thickness and collagen matrix integrity.

CONCLUSION

These findings demonstrate that reynosin exerts its potent anti-osteosarcoma activity by targeting the novel PRDX6-AKT-WEE1 signaling axis, suggesting its potential as a promising lead compound for the treatment of redox-sensitive malignancies.