Network Pharmacology and Molecular Docking Combined with Experimental Validation: The Mechanism of Brusatol in Regulating Glucose Metabolism in Lung Adenocarcinoma.

Abstract

Introduction: Lung cancer ranks first globally in cancer-associated mortality, with Lung Adenocarcinoma (LUAD) accounting for 35%-40% of all cases. Metabolic reprogramming is a hallmark of malignant cells, and aberrant glycolytic turnover acts as a major driver of LUAD progression. Natural small molecules have long attracted attention in anticancer research. Brusatol (Bru), an active ingredient extracted from the traditional Chinese medicinal plant Brucea javanica, exhibits robust antitumor activity. Nevertheless, its precise molecular mechanism of action in LUAD remains incompletely understood. To address this gap, the present study was designed.

Methods: We integrated network pharmacology and machine learning algorithms to screen candidate therapeutic targets of brusatol in LUAD and performed functional enrichment analyses. Molecular docking was further employed to identify core target genes. A panel of in vitro cellular assays was then conducted to evaluate the effects of brusatol on LUAD cell proliferation, migration, and invasion, as well as on cellular glucose uptake and the production of glycolytic end products (lactate and pyruvate). Western blotting was used to measure the expression of key proteins in the AKT/mTOR cascade and glucose metabolic machinery. Finally, we combined siRNA-mediated GPI silencing with brusatol treatment to validate the regulatory relationship between brusatol and GPI.

Results: GPI was confirmed as the core target of brusatol, and its high expression was closely correlated with poor prognosis in LUAD patients. Enrichment analyses revealed that GPI is primarily involved in metabolic pathways, glycolysis, and gluconeogenesis signaling. Cellular experiments demonstrated that brusatol suppressed LUAD cell proliferation, migration, and invasion in a dose-dependent manner. It also reduced glucose consumption and glycolytic metabolite production, while inhibiting AKT pathway activity and the expression of glucose metabolism-related proteins. Combined intervention further enhanced these inhibitory effects.

Discussion: Against the broad research backdrop of tumor metabolic reprogramming and natural productbased targeted therapy, this study uncovered the vital role of GPI as a key glycolytic hub in mediating LUAD malignancy. We also elucidated a novel molecular mechanism wherein brusatol reverses glycolytic dysfunction by targeting GPI to modulate the AKT/mTOR signaling axis. Previous investigations predominantly focused on the broad-spectrum anticancer efficacy of brusatol. In contrast, our research precisely delineates its core functional target and downstream signaling cascade from the perspective of metabolic regulation. This work fills detailed mechanistic gaps regarding how natural products intervene in glycolytic metabolism in LUAD and provides fresh insights into how small-molecule natural compounds remodel tumor metabolism.

Conclusion: Brusatol curbs glucose metabolic reprogramming by downregulating GPI expression, thereby arresting the malignant progression of LUAD. Interference with GPI function can further potentiate the antitumor activity of brusatol. Collectively, these findings offer a novel mechanistic rationale and preliminary in vitro evidence for glycolysis-targeted research in LUAD.