Mechanism of Banxia Xiexin Decoction in Preventing and Treating Gastric Cancer Based on Transcriptomics, Network Pharmacology and Experimental Validation

OBJECTIVE  To explore the mechanism of action of Banxia Xiexin decoction(BXD) in preventing and treating gastric cancer(GC) based on transcriptomics and network pharmacology, combined with experimental validation.

METHODS The active ingredient of BXD was quality controlled and identified by UPLC-Q-Orbitrap MS/MS technology, and the relevant targets of the drug ingredient were predicted by TCMSP, DrugBank and SwissTarget Prediction platforms. Genecards, OMIM, DisGeNet and other databases were used to identify GC genes, STRING databases were used to construct protein interaction networks, Metascape databases were used for KEGG and GO enrichment analysis, and cytoscape was used to construct active ingredient-target-pathway network diagrams. RNA-seq sequencing technology was used to screen the differentially expressed genes(DEGs) of GC cells MGC-803, and the coincident targets between RNA-seq and network pharmacology were analyzed, and the key targets of BXD for the treatment of GC were obtained, and the protein-protein interactions network(PPI), GO and KEGG enrichment analysis were performed using STRING and Metascape databases. CCK8, apoptosis assay and cell migration assay were used to detect the molecular function of cells, and qPCR was used to detect the mRNA expression of key targets.

RESULTS UPLC-Q-Orbitrap MS/MS combined with network pharmacology identified a total of 29 compounds and 859 potential targets in BXD. A total of 7695 GC-related genes were obtained from databases such as Genecards, OMIM, and DisGeNet, and 569 potential targets in BXD were identified for the treatment of GC. A total of 328 DEGs were detected by RNA-seq sequencing combined with network pharmacology, and the KEGG results showed that BXD might prevent GC through the PI3K-Akt signaling pathway, including PIK3CA, Akt1, EGFR, TNF and others were classified as hub genes, and in vitro cell experiments confirmed that BXD medicated serum and PI3K inhibitors-LY294002 could inhibit the proliferation of GC cells, promote apoptosis, inhibit the migration of GC cells, and play a role in reducing the expression of EGFR, PIK3CA and TNF in the PI3K-Akt signaling pathway in GC cells MGC803 cells.

CONCLUSION  BXD has the effect of inhibiting the proliferation of GC and delaying the development of GC. Its mechanism of action may be related to the regulation of PI3K-Akt signaling pathway and the alleviation of inflammatory response.