Exploration of the role and mechanism of Rhizoma Paridis total saponins in osteosarcoma based on SPI1/LCN2-mediated ferroptosis.

Background: Osteosarcoma (OS) is a highly aggressive bone malignancy with limited therapeutic options and poor prognosis, particularly in cases of recurrence or metastasis. Recent studies have identified ferroptosis as a promising therapeutic target in OS, with the SPI1/LCN2 regulatory axis emerging as a critical modulator of this process. We hypothesized that Rhizoma Paridis total saponins (RPTS) exert anti-osteosarcoma effects by inducing ferroptosis through inhibition of the SPI1/LCN2 axis.

Methods: assessments using OS cell lines MG-63 and Saos-2 included cell counting kit-8 (CCK-8) assays for viability, colony formation for proliferation, scratch wound healing for migration, and Transwell chambers for invasion. Ferroptosis markers were evaluated using colorimetric Fe detection, flow cytometric apoptosis analysis, DCFH-DA probes for reactive oxygen species (ROS), DTNB colorimetry for glutathione (GSH) levels, and western blot (WB) for SLC7A11 and GPX4 expression. A subcutaneous xenograft mouse model received OS cell injections for validation of tumor growth parameters and protein expression. Bioinformatics analysis was conducted to screen OS-specific differentially expressed genes, followed by validation in both and experiments using RT-qPCR and WB.

Results: RPTS significantly inhibited OS cell viability, proliferation, migration, and invasion capacity while promoting Fe accumulation and ROS generation. , RPTS treatment significantly inhibited tumor growth . Bioinformatics analysis identified LCN2 as the core ferroptosis mediator regulated by upstream transcription factor SPI1. RPTS downregulated SPI1 and LCN2 expression and . Both LCN2 overexpression and SPI1 activation reversed RPTS-mediated ferroptosis induction. SPI1 overexpression with LCN2 knockdown attenuated the promoting effect of RPTS on ferroptosis.

Conclusion: RPTS triggers ferroptosis-mediated OS suppression through SPI1/LCN2 axis inhibition, providing a novel therapeutic strategy to improve clinical outcomes in OS management.