Phenolic Acid Composition and Apoptosis-Inducing Activity of Rhynchosia nulubilis Extracts Fermented with Ganoderma lucidum

This study investigated the bioactive properties of fractions derived from Rhynchosia nulubilis cultivated with Ganoderma lucidum mycelium (RNGM), focusing on cytotoxic and apoptosis-related responses in cancer cells. Fractions obtained using n-hexane, chloroform, ethyl acetate, and water were evaluated for cytotoxic effects against A549, Hep3B, HeLa, and HeLa229 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The phenolic acid composition of each fraction was determined by high-performance liquid chromatography (HPLC). Among the fractions, the ethyl acetate fraction showed the highest total phenolic acid content and exhibited the strongest cytotoxic activity, particularly against HeLa cells. Apoptosis induction was supported by increased caspase-3/7 activity, apoptotic nuclear morphology observed by 4′,6-diamidino-2-phenylindole (DAPI) staining, and accumulation of cells in the sub-G1 phase. In addition, treatment with the ethyl acetate fraction upregulated p53 and Bax mRNA expression and increased the Bax/Bcl-2 ratio. These findings suggest that the ethyl acetate fraction of RNGM induces apoptosis-mediated growth inhibition in cervical cancer cells.