Steroidal glycosides from Mallotus philippensis induce apoptosis in MCF-7 breast cancer cells via MTT and DAPI assays

Abstract. Manikyam HK, Joshi SK, Patil SB, Vakadi S, Patil AR. 2025. Steroidal glycosides from Mallotus philippensis induce apoptosis in MCF-7 breast cancer cells via MTT and DAPI assays. Asian J Nat Prod Biochem 23: 47-51. Mallotus philippensis (Lam.) Müll.Arg., commonly referred to as the kamala tree, is a widely known ethnomedicinal plant possessing diverse pharmacological activities. Despite its traditional applications, modern research into its anticancer properties remains underexplored. The present study investigates the in vitro anticancer activity of steroidal glycoside-enriched fractions derived from the aerial parts of M. philippensis against the MCF-7 breast cancer cell line. Ethanolic extraction followed by acetone precipitation was employed to obtain the glycoside-rich fraction. Phytochemical screening confirmed the presence of steroidal glycosides using Liebermann–Burchard and Salkowski tests. To evaluate cytotoxicity and apoptosis induction, MTT cell viability assay and DAPI nuclear staining assay were performed. The MTT assay results revealed a concentration-dependent inhibition of cell growth, achieving complete (100%) growth inhibition at 200 µg/mL. DAPI staining further confirmed the pro-apoptotic activity of the extract, showing prominent nuclear fragmentation and chromatin condensation, with an apoptotic index of 67.87 ± 6.13%. These effects were comparable to the standard positive control, sodium Lauryl Sulfate (SLS), suggesting a potent anticancer potential. The findings support the hypothesis that steroidal glycosides present in M. philippensis can significantly induce apoptosis in hormone-dependent breast cancer cells. The anticancer effect is likely mediated through mitochondrial apoptotic pathways, characteristic of steroidal compounds. This study provides a strong rationale for further investigation into the bioactive constituents of M. philippensis, including isolation, molecular mechanism elucidation, and in vivo validation.