Mechanistic evaluation of pachymic acid in breast cancer via network pharmacology and in vitro validation

Breast cancer(BC) is a major cause of cancer-related mortality in women worldwide, with current therapies often limited by toxicity and drug resistance. Pachymic acid (PA), a natural compound, demonstrates antitumor potential with relatively low systemic toxicity. However, its mechanism in triple-negative breast cancer (TNBC) remains unclear.

This study aims to deeply explore the anti-tumor effect of PA on TNBC. The intersection targets of PA and BC were screened through multiple databases, and the key genes were determined through TCGA survival analysis. The potential mechanism was simulated and studied through methods such as network pharmacology, molecular docking and molecular dynamics. Cell proliferation ability was evaluated by CCK-8 assay and colony formation assay. Cell migration and invasion abilities were detected by scratch assay and Transwell assay respectively. Gene expression levels were analyzed by qPCR and Western Blot, and the degree of lipid peroxidation was evaluated by malondialdehyde (MDA) assay.

Through multi-database screening and TCGA survival analysis, PTGS2, NOS2 and ABCB1 were identified as key targets. The TNBC cell line MDA-MB-231 was used as the experimental model. PA significantly inhibited the proliferation, migration and invasion of MDA-MB-231 cells. Mechanistic studies demonstrated that PA concurrently upregulates PTGS2 and NOS2 expression while downregulating ABCB1. Notably, the upregulation of PTGS2 was significantly correlated with increased lipid peroxidation, suggesting its crucial role in ferroptosis. The coordinated upregulation of NOS2 and downregulation of ABCB1 further enhanced cellular sensitivity to ferroptosis.

This study is the first to reveal that PA induces ferroptosis in TNBC cells by regulating the PTGS2/NOS2/ABCB1 signaling axis, providing new potential targets for TNBC treatment. However, this study has limitations, as validation has been conducted only at the cellular level and not yet confirmed in animal models or clinical samples. Future research should validate this mechanism in more TNBC models and across different molecular subtypes of breast cancer to promote the clinical application of PA.