Anticancer activity of Hybanthus enneaspermus (Linn) F. Muell extracts on A549-CCL185 (Lung Adenocarcinoma cell)

Aim: This study aims to elucidate the fundamental apoptotic mechanisms and evaluate the anticancer efficacy of whole plant extracts of Hybanthus enneaspermus (Linn.) F. Muell against human lung adenocarcinoma (A549-CCL185) cells. Methods: The whole plant was gathered, analyzed, shade-dried, and then extracted in stages with water, ethanol, ethyl acetate, and chloroform using the Soxhlet and cold maceration techniques. We used the MTT assay to find out how toxic the extracts were to A549-CCL185 cells. To verify apoptosis, DNA fragmentation was analyzed through agarose gel electrophoresis, and morphological alterations were assessed using acridine orange/ethidium bromide (AO/EtBr) dual staining. Results: The ethanolic extract (EEHE) demonstrated the most significant cytotoxic effect among all tested extracts (IC₅₀ = 37.88µg/ml), while the ethyl acetate extract (EAEHE) displayed moderate activity (IC₅₀ = 215.39 µg/ml). AO/EtBr staining showed clear signs of apoptosis, such as breaking up of chromatin, condensation of the nucleus, and shrinking of the cell. In cells treated with EEHE, apoptotic cell death was verified by DNA ladder formation. Phenolic and flavonoid compounds that encourage mitochondrial dysfunction brought on by reactive oxygen species (ROS) and cause programmed cell death are thought to be responsible for the cytotoxicity. Conclusion: Hybanthus enneaspermus's ethanolic extract significantly inhibits the growth of A549-CCL185 lung adenocarcinoma cells and induces apoptosis. These results point to its potential as a natural source of chemotherapeutic agents and validate its use in ethnomedicine. It is necessary to conduct additional research that includes in vivo validation and the isolation of active ingredients.