Fluorescent Staining–Assisted Evaluation of the Anticancer Potential of Nyctanthes arbor‐tristis Extracts in HepG2 Liver Cancer Cells

ABSTRACT The current study's initial goal was to investigate the anticancer activity of the extracts from the flowering plant Nyctanthes arbor‐tristis. The ethanol extracts' phytochemical analysis was qualitatively determined. It was determined that the various phytochemical substances had antibacterial, antioxidant, and anticancer effects. GC‐MS analysis revealed compounds like maleic anhydride, butanedioic acid, and oxalic acid emphasize the antioxidant and anticancer properties. MTT assay was investigated against three different cell lines (HepG2, A549, and MCF7). High level of resistance to HepG2 liver cancer cell lines was noted with ethanol flower extracts with IC50 value of 67.98 μg/mL. EtBr staining assay revealed HepG2 apoptotic cells displaying nuclear and membrane alterations when exposed to 70 μg/mL of ethanol extract. The cells treated with 70 μg/mL of extract showed a reduction of mitochondrial membrane potential, as evidenced by the MMP assay, which showed red fluorescence decreasing and green fluorescence increasing. DNA fragmentation analysis showed HepG2 cancer cells showed no signs of DNA fragmentation after being treated with 50 μM treatments of flower extracts for 72 h. Quantitative RT‐PCR analysis showed increased Bcl2 gene expression with fold variation level of about 0.086094579. The obtained results emphasized the significance of Nyctanthes arbor‐tristis extracts with a suggestion of chemotherapeutic medication to treat various cancers in the future. The study examines the phytochemical analysis of Nyctanthes arbor‐tristis extract. The extract is also studied for their effective anticancer activity using various assays and staining techniques that paves a way for their therapeutic value.