To screen and identify the potential targets and pathways of Danthron against breast cancer based on network pharmacology, and to verify its mechanisms through transcriptomic analysis and in vitro and in vivo experiments.

Multiple databases were integrated to screen the common targets of Danthron and breast cancer for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. CCK-8 and colony formation assays were used to evaluate cell proliferation. Transcriptome sequencing and quantitative real-time polymerase chain reaction (qPCR) were performed to analyze the expression of Cxcl10, Isg15, IL-6, Nlrp3, and Bax and other related genes. A mouse 4T1 xenograft model was established to observe and assess the antitumor effect of Danthron.

A total of 84 intersecting targets were obtained, mainly enriched in the Apoptosis, p53, PI3K-Akt, NOD-like receptor, and Cytokine signaling pathways. Danthron significantly inhibited breast cancer cell proliferation (IC₅₀=30.05 μmol·L^‐1), upregulated interferon-stimulated genes (Irf7, Isg15, Ifit1) and chemokines (Cxcl10, IL‐6), and upregulated the expression of Nlrp3 and Bax. In vivo, Danthron significantly suppressed 4T1 xenograft growth and reduced Ki-67 expression.

Danthron exerts antitumor activity against breast cancer by activating interferon responses and inflammation-apoptosis-related pathways, providing a theoretical basis for its development as a potential natural anticancer agent.