Integrating Network Pharmacology and in Vitro Experiments to Explore the Potential Mechanisms of Dianthus Caryophyllus Methanolic Extract in Inhibiting the Proliferation of Triple-Negative Breast Cancer Cells
KANG Shiyao1#, DENG Xuming1#, LI Huimin1, TANG Yuheng1, LI Ming1, TANG Wenru1, ZHOU Xuhong2*, SHENG Miaomiao1*
TNBC (triple-negative breast cancer), characterized by the absence of ER (estrogen receptor), PR (progesterone receptor), and human HER2 (epidermal growth factor receptor 2) expression, exhibits high aggressiveness, limited therapeutic options, and poor prognosis. Although Dianthus caryophyllus L. possesses antican cer potential, its anti-TNBC mechanisms remain incompletely elucidated. This study integrated network pharmacol ogy, molecular docking, and in vitro validation to investigate the mechanism of carnation methanolic extract against TNBC. A total of 883 secondary metabolites were identified in carnation using HPLC-MS, and 140 bioactive components were selected through the CTD (comparative toxicogenomics database). By intersecting TNBC-related targets from GeneCards and OMIM databases, 82 potential therapeutic targets were identified. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analyses revealed that oxidative stress levels and the PI3K/AKT and FoxO signaling pathways were key regulatory axes for the anticancer effects of carnation against TNBC. Based on the STRING database, a PPI network was constructed, 10 core target genes were screened out, and a prognostic risk model was established. Molecular docking showed that the main bioactive components of carnation (e.g., betulinic acid and kaempferol) have significant binding activity with the core target genes. Further in vitro experiments demonstrated that the carnation methanolic extract significantly inhibited the proliferation of MDA-MB-231 and MDA-MB-468 cells in a concentration- and time-dependent manner. Additionally, it signifi cantly increased intracellular ROS levels and downregulated the expression of key proteins in the PI3K/AKT and FoxO signaling pathways. These results are consistent with the predictions from network pharmacology and mo lecular docking analyses. In summary, through a multidimensional validation of “components-targets-pathways experiments,” this study elucidated that the carnation methanolic extract exerts anticancer effects against TNBC by modulating oxidative stress and the PI3K/AKT-FoxO signaling axis. These findings provide experimental evidence for the development of carnation as a potential adjuvant therapeutic agent for TNBC.
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