Matrine as a potent inhibitor of canine mammary tumors: Insights from network pharmacology, molecular docking, molecular dynamics and in vitro analysis.

Abstract

Canine mammary tumor (CMT) is the most prevailing neoplasms in female dogs. Matrine demonstrates anti-tumor effects in various organs, its mechanism in CMT treatment remains unclear. This study involved in network pharmacology, molecular docking, dynamics simulations, cell viability, inhibations and migration, to clarify matrine's therapeutic action against CMT. Potential gene/protein targets were identified through PubChem (Matrine), and GeneCards (CMT), with point of intersection targets analyzed via STRING PPI network and Cytoscape. Top 10 hub genes were selected for gene ontology/KEGG pathway analyses (p&#x202f;<&#x202f;0.05). Molecular docking (JAK2, AR, HDAC2, HDAC6, and NR3C1) revealed strong matrine-JAK2 binding (-7.8&#x202f;kcal/mol), and dynamics simulations were confirmed by molecular dynamics simulations. Pathway analysis implicated JAK-STAT and PI3K-Akt signaling in matrine's as anti-tumor effects. In vitro results showing that the maximum noncytotoxic concentrations of matrine of canine primary mammary epithelial cells (cPMECs), where the cell viability remained above 90&#x202f;% at concentrations 280&#x202f;&#xb5;M and &#x2264;&#x202f;560&#x202f;&#xb5;M, respectively, indicating this as the maximum safe concentration for cPMECs, and had a proliferation inhibitory effect time-dependently (12, 24, 48 hrs) on CHMm and CHMp cells within a safe concentration range, and suppression of CMT cells via CCK-8 assays. In 12&#x202f;h, moderate inhibition was detected, which increased at 24&#x202f;h, and was most prominent at 48&#x202f;h. The migration ability of cells decreased at 24&#x202f;h, respectively. Notably, the 560&#x202f;&#xb5;M concentration resulted in over 50&#x202f;% inhibition in both cell lines after 48&#x202f;h. Future research should investigate in vivo efficacy to progression matrine as a veterinary oncotherapeutic.