The Tanshinones of Salvia miltiorrhiza Bunge Suppress GES-1 Apoptosis via Inhibiting EGFR-STAT3 Signaling Pathway in vitro

Aim: Chronic atrophic gastritis (CAG), a precancerous lesion of gastric cancer. Current therapies face challenges including drug resistance and incomplete efficacy. Tanshinones exhibit anti-inflammatory and anti-tumor properties, yet their role in CAG remains unexplored. The present study aimed to explore the therapeutic effect and underlying mechanism of tanshinones on N-methyl-N’-nitro-N-nitrosoguanidine (MNNG)-induced CAG in vitro. Methods: The active tanshinones were screened from CNKI, PubMed and TCMSP databases. The CAG-related targets were collected from GeneCards database. Cytoscape 3.9.0 software was used to construct a compound-target network. A protein-protein interaction (PPI) network was constructed using a string database. The potential mechanism was analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Molecular docking was completed by Autodock software. An in vitro CAG model was established using the human gastric epithelial cell line GES-1, induced by MNNG. Cell viability of GES-1 was measured by Cell Counting Kit-8 (CCK-8) assay. TUNEL assay was used to evaluate cell apoptosis and protein expression of Epidermal growth factor receptor (EGFR), Signal transducer and activator of transcription 3 (STAT3) etc was determined by Western blot. Results: The compound-target network analysis found 12 active tanshinones and 42 common targets. The key targets were Cysteinyl Aspartate-specific Proteinase-3 (Caspase-3 (CASP3)), EGFR, STAT3, PTGS2, MMP2, ERBB2, and PPARG. Through enrichment analysis, 656 GO terms were enriched, whereas the top 15 pathways, including proteoglycans in cancer and apoptosis, were identified. Through molecular docking, cryptotanshinone, tanshinone IIA, dihydrotanshinone I, and tanshinone I were found to be stably bound to EGFR and STAT3. In vitro experiments indicated that tanshinones could significantly inhibit MNNG-induced GES-1 cell apoptosis, and markedly lower the protein expression of EGFR, STAT3, and Caspase-3. Conclusion: Tanshinones combination exhibited a positive effect on anti-CAG induced by MNNG in vitro, and this function is correlated with the suppression of EGFR-STAT3 signaling pathway and cell apoptosis.