Astragaloside IV potentiates cisplatin sensitivity in triple-negative breast cancer via STING signaling pathway activation

Background: Astragaloside IV (As), a bioactive tetracyclic triterpenoid saponin from Astragalus membranaceus, exhibits diverse pharmacological properties including anti-inflammatory, Anticancer and immunomodulatory activities. However, its potential role as a targeted therapeutic agent for cisplatin sensitization in triple-negative breast Cancer (TNBC) via the STING signaling pathway remains unexplored.

Purpose: This study aimed to investigate the cisplatin-sensitizing potential of As in TNBC and elucidate its underlying molecular mechanisms.

Methods: Bioinformatic analysis identified key module genes from public databases using weighted gene co-expression network analysis (WGCNA), followed by feature selection via least absolute shrinkage and selection operator (LASSO) regression. Immune microenvironment characterization was performed using CIBERSORT deconvolution analysis. Potential targets of As in TNBC were systematically identified through multi-database mining, protein-protein interaction (PPI) network construction, pathway enrichment analysis and molecular docking simulations. Transcriptomic profiling revealed treatment-induced differentially expressed genes and enriched signaling pathways in murine tissues. The combinatorial anti-tumor efficacy was evaluated using orthotopic TNBC model (4T1 cell-derived) in vivo and 4T1, MDA-MB-231, and MCF-7 cell lines in vitro. Mechanistic validation was conducted through integrated in vivo and in vitro experiments.

Results: Integrated bioinformatics analysis (WGCNA, LASSO, and CIBERSORT) identified STING within a chemotherapy-sensitive module among six endoplasmic reticulum (ER)-associated gene co-expression networks. STING exhibited significant enrichment in antitumor immune pathways and correlated with heterogeneous immune cell infiltration. KEGG pathway analysis implicated immune/inflammatory signaling and DNA damage responses in the cisplatin-sensitizing effects of As. Molecular docking confirmed stable binding between As and STING (binding energy: -36.09 kcal/mol). In vivo, As synergistically enhanced cisplatin's tumor growth inhibition while mitigating cisplatin-induced toxicity, including weight loss, nephrotoxicity, and systemic inflammation. Transcriptomic and immunohistochemical analyses validated As-mediated DNA damage, STING pathway activation, immune cell recruitment. In vitro mechanistic studies using DNase I and STING inhibitor (C-176) demonstrated As potentiated cisplatin via cell cycle arrest, mitochondrial dysfunction and STING cascade activation. C-176 weakened the antitumor activity of As- cisplatin combination treatment.

Conclusion: As synergistically enhances the therapeutic efficacy of cisplatin against TNBC while ameliorating its adverse effects, primarily through activation of the STING signaling pathway and potentiation of antitumor immunity. This dual mechanism-simultaneously boosting host defense ("Fuzheng") and inhibiting tumor progression ("Kang`ai")-provides a molecular basis for the observed "Qi-tonifying" properties of As in traditional Chinese medicine (TCM), bridging TCM with contemporary immunopharmacology.

Astragaloside IV; Cisplatin; STING; Triple- negative breast cancer.