Exploring the Potential Mechanism of Ethyl Acetate Extract from Tumor Vine in Combination with Cisplatin for Treating Non-Small Cell Lung Cancer Based on Network Pharmacology and Cellular Experiments.

To investigate the mechanism of the ethyl acetate extract of Tumor Vine (PT-EAE) in combination with cisplatin (DDP) for the treatment of non-small cell lung cancer (NSCLC), using network pharmacology prediction and validation through cellular experiments. The chemical components of PT-EAE were identified using Ultra-Performance Liquid Chromatography-Quadrupole-Orbitrap Mass Spectrometry (UPLC-Q-Exactive-Orbitrap-MS). Network pharmacology was employed to predict the potential active components of PT-EAE and their molecular targets affecting NSCLC cell functions. A "PT-EAE-active components-targets-disease" interaction network was constructed. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on the potential targets to identify core targets and signaling pathways closely related to the active components of PT-EAE. Subsequently, in A549 cells, the effects of PT-EAE alone, DDP alone, and their combination (PT-EAE & DDP) were evaluated. Cell Counting Kit-8 (CCK-8) assay, wound healing assay, transwell assay, and flow cytometry were used to detect cell proliferation, migration, invasion, and apoptosis, respectively. Finally, western blotting and quantitative reverse transcription polymerase chain reaction (RT-qPCR) were utilized to detect the expression levels and phosphorylation status of the predicted core molecular proteins following treatment with PT-EAE and DDP. PT-EAE inhibited the proliferation, migration, and invasion of A549 cells, while promoting apoptosis. Compared to the DDP monotherapy group, the PT-EAE & DDP combination group reduced the IC50 of DDP for inhibiting A549 cell proliferation. Network pharmacology analysis identified 12 active components in PT-EAE, which were mapped to 124 NSCLC-related targets and multiple key signaling pathways. Among these, EGFR, Src, and PI3K were identified as potential core nodes. At the molecular level, PT-EAE downregulated the protein expression levels of EGFR and Src in A549 cells, as well as the phosphorylation levels of downstream molecules in the Src signaling pathway-PI3K, Akt, and mTOR. It also suppressed the mRNA expression of EGFR, Src, PI3K, Akt, and mTOR. The ethyl acetate extract of Tumor Vine, when combined with cisplatin, shows promising anti-tumor activity through multi-component, multi-target, and multi-pathway actions, with the PI3K/Akt/mTOR signaling pathway likely representing one of its therapeutic routes.