Phytochemical profiling and anticancer activity of the n-butanol fraction from Ardisia villosa extract: Inhibition of gastric cancer cell proliferation via cell cycle arrest and senescence induction

Medicinal plants serve as valuable sources for anticancer drug discovery. This study investigated the anticancer potential of the n-butanol fraction from Ardisia villosa extract against breast and gastric cancer cell lines. Phytochemical profiling using UPLC-QToF-MS in both positive and negative ESI modes identified 118 putative compounds, including flavonoids, lignans, alkaloids, triterpenoids, steroids, coumarins, and phenolic acids. The n-butanol fraction exhibited dose-dependent antiproliferative effects, with IC50 values of 60.2 µg/mL (MCF-7), 85.2 µg/mL (MKN45), and 51.7 µg/mL (AGS). In AGS gastric cancer cells, the extract significantly inhibited 3D tumorsphere formation and suppressed cell migration at concentrations as low as 50 µg/mL (p < 0.001). Additionally, n-butanol fraction extract markedly induced cellular senescence (p < 0.01). Mechanistic investigations revealed that the extract induced G0/G1 phase arrest by downregulating critical cell cycle regulators, including CCND1, CCNE1, CDK2, CDK3, CDK6, CDK8, and CDK9, while upregulating tumor suppressor and senescence-related genes such as p21, p53, p16, and p27 (p < 0.01). Molecular docking analyses further supported these findings by demonstrating strong binding affinities of phytochemicals to key cell cycle regulatory proteins, suggesting a direct molecular basis for their antiproliferative effects. In conclusion, the n-butanol fraction of Ardisia villosa displays potent anticancer activity, particularly in gastric cancer cells, through multi-targeted mechanisms involving cell cycle inhibition and senescence induction, and holds promise as a natural source for future anticancer therapeutics.