Integrated Detection and Validation of the Mechanistic Basis for Radix Polygoni cynanchoidis Activity in Anaplastic Thyroid Cancer

Background Radix Polygoni cynanchoidis is a long-standing traditional remedy historically used to manage pulmonary tuberculosis, persistent cough, rheumatism, and other chronic conditions. While earlier studies have mainly characterized its phytochemistry, no investigations employing advanced mass spectrometry have yet been published. Methods This study was developed to systematically profile the chemical constituents of Radix P. cynanchoidis and to clarify its pharmacological mechanisms in anaplastic thyroid cancer (ATC) using a network pharmacology approach. These predictions were then substantiated by cell-based assays and Western blotting of key compounds. Chemical characterization was carried out using UPLC-Q-Exactive Orbitrap/MS, with compounds assigned based on diagnostic fragment ions, fragmentation pathways, and cross-referencing with ChemSpider and in-house libraries. Predicted molecular targets and pathways were mapped via network pharmacology. Anti-proliferative effects in ATC cells were assessed by CCK-8 assay, while apoptosis was measured using flow cytometry and western blotting. Results Sixteen anthraquinones were identified in analyses of Radix P. cynanchoidis. Network pharmacology revealed 735 Gene Ontology biological processes and 142 KEGG pathways intersecting with ATC-related targets. In vitro assays demonstrated that principal constituents significantly inhibited ATC cell proliferation and enhanced apoptotic activity. Western blotting further confirmed an increase in Caspase-3 expression following treatment. These findings indicate that Radix P. cynanchoidis exerts its anti-ATC effects via multiple bioactive compounds acting on a wide range of targets and pathways. Conclusions This study provides the first mass spectrometry-based compositional analysis and preliminary mechanistic evidence for the therapeutic potential of this herbal preparation.