Network pharmacology and chemoproteomics unveil the mechanism of Total Flavonoid Aglycones Extract inhibiting EMT in A549 cell line

Non-small cell lung cancer (NSCLC) is a worldwide disease with high morbidity and mortality. The epithelial-mesenchymal transition (EMT) process promotes cancer cell migration and invasion, leading to NSCLC metastasis. Total Flavonoid Aglycones Extract (TFAE) isolated from Scutellaria baicalensis was reported to inhibit tumor growth and induce apoptosis. In this study, combining network pharmacology and stable isotope dimethyl-labeled proteomics technology, we found the inhibitory effect of TFAE on the EMT process of A549 cells, and established the relationship between TFAE, PI3K/Akt signaling pathways, TWIST1, and glycolysis pathway. LY294002 was incubated with A549 cells to inhibit PI3K/Akt signaling pathway, and small interfering RNA (siRNA) method was applied to knock down the TWIST1 gene in A549 cell. Compared with normal cells, LY294002-treated/ TFAE-treated/ TWIST1-silenced / TFAE and siTWIST1-treated cells showed poor migration and invasion capabilities, increased expression of epithelial markers and decreased expression of mesenchymal markers. All the results suggested that TFAE inhibits PI3K/Akt pathway and TWIST1 and then inhibits glycolysis pathway, thereby inhibiting the EMT process of A549 cells.