Investigating Anticancer Properties of Artemisia annua in Human Pancreatic Cancer Cells: Insights from Network Pharmacology, Molecular Docking, Dynamics Simulations, and Experimental Testing

This study examines the bioactive compounds of Artemisia annua (A. annua) and their therapeutic potential in treating Prostate Cancer (PC) through network pharmacology, bioinformatics, and in vitro validation. It screened 126 bioactive principles from the TCMSP database and then reduced them to 22 based on drug-likeness, oral bioavailability, and permeability. The subsequent screening yielded 396 biological targets, with possible duplicates eliminated, and further reduced to 72 putative targets. The comparative analysis with 14,146 PC-related genes resulted in the identification of 62 common targets. Network analysis highlighted three major compounds, Kaempferol, Sitosterol, and Quercetin, which interact with these targets. Molecular docking indicated strong binding affinities of Kaempferol to key hub genes EGFR, MMP9, and GSK3B (Vina scores: -8.4, -9.0, -7.7) that were further supported by stable interactions in molecular dynamics simulations. In vitro studies indicated that the ethanol-DCM extract of A. annua exhibited statistically significant cytotoxic effects on PANC-1 cells. Cell viability assays showed dose- and time-dependent inhibition after 48 hours of treatment. Migration and invasion assays showed a significant reduction in cell movement across transwell membranes at 60 µg/mL (p < 0.01) and 120 µg/mL (p < 0.001). Invasiveness was also significantly suppressed at 60 µg/mL (p < 0.01) and 120 µg/mL (p < 0.001), indicating anti-metastatic potential. Flow cytometric cell cycle analysis showed significant G2/M-phase arrest and 45.6% of cells arrested in G2/M-phase at 120 µg/mL compared to 18.4% in the untreated controls were found (p < 0.001). Annexin V-FITC/PI staining evidenced apoptosis induction, with 28.3% apoptotic cells at 120 µg/mL levels compared to 6.1% in controls (p < 0.001). These findings demonstrate the potential of A. annua to modulate critical molecular pathways in PC, offering promising insights for developing novel therapeutic strategies.