Ardisiaoside A, a new triterpenoid glycoside from Ardisia gigantifolia, induces cell senescence and targets cancer stem cells in gastric cancer.

This study aimed to isolate bioactive constituents from Ardisia gigantifolia and assess their effects on gastric cancer stem cells (CSCs) properties. Cytotoxicity and proliferation were quantified using the MTT assay; cell-cycle distribution, CD44 expression, and apoptosis/senescence markers were evaluated by flow cytometry and β-galactosidase staining; reactive oxygen species (ROS) generation and tumorsphere formation were analyzed by fluorescence microscopy and 3D culture; gene expression was measured by qPCR; molecular docking probed interactions with CSCs markers and Notch signaling proteins. Four compounds were isolated, including Ardisiaoside A, β-sitosterol, and two long-chain n-alcohols (octacosan-1-ol, nonacosan-1-ol). Ardisiaoside A was reported from this species for the first time and showed the greatest potency, reducing viability at low micromolar doses (IC: 1.08 ± 0.37 µM in AGS, 2.50 ± 0.72 µM in MKN45 cells). It suppressed migration, induced G2/M arrest, and triggered senescence. CD44 surface levels was markedly reduced, and tumorsphere number and size were inhibited at 2.5 µM. qPCR and immunofluorescence demonstrated notable downregulation of stemness-related genes (OCT4, NANOG) and Notch pathway components, consistent with reduced self-renewal. Molecular docking supported the binding of Ardisiaoside A to NANOG, OCT4, CD44, and Notch proteins (NOTCH1, HES1, DLL1, DLL4), consistent with target inhibition. In conclusion, Ardisiaoside A, a newly identified triterpenoid glycoside from Ardisia gigantifolia, represents a promising candidate that inhibits cell proliferation by reducing cancer stem-cell populations and inducing cellular senescence in gastric cancer.