Tongguanteng Injection Induces Apoptosis of Colorectal Cancer Cells Via Inhibiting PTGS2 Mediated PD-L1 Checkpoint Pathway: Insights from Network Pharmacology and Experimental Validation.

This study was designed to systematically assess the active ingredients of Tongguanteng injection (TGT) and its molecular mechanisms in CRC, with further experimental validation of the implicated pathways. The active ingredients of TGT were obtained from the Herb database. The SwissTargetPrediction database was used to predict the potential targets of these active ingredients. Targets of CRC were obtained from GeneCards, OMIM, DisGeNET, and TTD databases. Cytoscape 3.9.1 was used to build ingredient-target-disease and PPI networks, followed by GO and KEGG enrichment analyses. The interaction between active ingredients and receptor proteins was analyzed through molecular docking using Autodock Tools 1.5.7. Bioinformatics validation of core targets was carried out with UALCAN, KM-plotter, HPA, TIMER, and cBioPortal. Furthermore, in vitro studies were performed to verify the therapeutic efficacy of TGT and its active constituent Tenacissoside G (TG) against CRC and to confirm the critical mechanisms. Network analysis revealed 54 TGT ingredients and 177 CRC-related targets, among which AKT1, EGFR, BCL2, and PTGS2 emerged as the key core targets. KEGG pathway analysis revealed significant involvement of the PD-L1/PD-1 checkpoint pathway. Molecular docking analysis showed high-affinity binding of the active compounds TG to PTGS2. Bioinformatics analysis verified differential expression patterns of PTGS2, EGFR and BCL2 relating to prognostic significance between colorectal cancer and normal tissues. In vitro validation demonstrated TGT and its active ingredients TG induce CRC cell apoptosis by suppressing PTGS2-mediated PD-L1 signaling. TGT and TG play an anti-CRC role by targeting PTGS2 and inhibiting PD-L1/PD-1 to promote apoptosis. This study indicates the therapeutic potential of TGT in CRC.