Integrating Network Pharmacology, Transcriptome Analysis, and In Vitro Experiment to Investigate Molecular Mechanism...

Liensinine (LIE), a bioactive alkaloid from Nelumbo nucifera, exhibits proapoptotic and anticancer properties. Its effect and mechanism in pancreatic cancer remain unexplored, representing a novel research direction. We employed an integrated strategy combining network pharmacology, transcriptome sequencing, and experimental validation to investigate LIE's action against PANC-1 pancreatic cancer cells. Key targets and pathways were identified via bioinformatics analyses. Molecular docking and survival analysis were conducted to verify core targets. In vitro assays including CCK-8, lactate dehydrogenase release, Hoechst/PI staining, flow cytometry, wound healing, and transwell invasion were used to assess cytotoxicity, apoptosis, migration, and invasion. Western blotting validated protein-level changes. Network pharmacology revealed 90 overlapping targets of LIE and pancreatic cancer, with the Phosphatidylinositol 3-k (PI3K)/Serine/Threonine Kinase (AKT) pathway being significantly enriched. Molecular docking indicated a strong binding affinity of LIE to Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha (PIK3CA) and AKT1. Survival analysis associated high PIK3CA expression with poorer prognosis in pancreatic cancer patients. In vitro, LIE dose-dependently inhibited PANC-1 cell proliferation, migration, and invasion while significantly inducing apoptosis. Western blotting confirmed that LIE treatment effectively suppressed the activation of the PI3K/AKT signaling pathway. Our study is the first to demonstrate that LIE exerts potent antitumor effects against pancreatic cancer cells by inducing apoptosis and inhibiting malignant phenotypes through suppression of the PI3K/AKT pathway. These findings highlight LIE as a promising novel therapeutic candidate for pancreatic cancer treatment.