Aquilaria Crassna Pierre Ex Lecomte Leaf Extract Induces Apoptosis and Immunogenic Cell Death in Breast Cancer Cells, Enhancing Anti-Tumor Effects of Dendritic Cells

Abstract

Aquilaria crassna (A. crassna) Pierre ex Lecomte, commonly known as agarwood, is traditionally used in medicinal preparations. Previous studies suggest anti-cancer properties in A. crassna leaf extract but the mechanisms and its involvement in immunogenic cell death (ICD) remain unexplored. This study investigates the impact of A. crassna Pierre ex Lecomte leaf extract (AE) on anti-cancer activity, specifically inducing ICD in triple-negative breast cancer (TNBC), and enhancing dendritic cell (DC) anti-tumor effects. A. crassna water extract underwent HPLC analysis. TNBC cells (MDA-MB-231) and non-tumorigenic epithelial cell lines, fibrocystic disease, (MCF-10A) were treated with different concentrations of AE for cytotoxicity testing by MTT assay. To assess ICD induction, danger-associated molecular patterns (DAMPs) including ectoCRT, secreted ATP, and HMGB1 were measured through staining, ATP bioluminescence, immunoblotting, and ELISA, respectively. AE-treated cells and DAMP-containing supernatants were administered to monocyte-derived DCs, evaluating impact on DC immunophenotype, maturation, and phagocytosis via flow cytometry. HPLC analysis identified AE compounds: iriflophenone 3,5-C-β-D-diglucoside, iriflophenone 3-C-β-D- glucoside, mangiferin, and genkwanin 5-O-β-primevoside (concentrations: 4.80, 1.04, 4.54 and 0.18 %w/w). MDA-MB- 231 treated with AE exhibited ICD induction, evident in increased ectoCRT, secreted ATP, and HMGB1 levels. DCs exposed to AE-treated cells and DAMP-containing supernatant displayed enhanced phagocytic activity and maturation, with elevated CD86, CD80, CD83, and HLA-DR expression. In conclusion, AE exhibits immunomodulatory potential by inducing ICD in TNBC, suggesting therapeutic applications for its anti-cancer effects and a promising role in cancer immunotherapy.