Cytotoxic profiling of Hedera pastuchovii in human cancer cell lines and its genoprotective effects on normal lymphocytes.

The genus Hedera L. has long been recognized as a valuable source of anticancer agents, such as triterpenoid saponins. In contrast, Hedera pastuchovii Woronow remains largely unexplored. This study investigates the cytotoxic profiles and genoprotective potential of H. pastuchovii leaf extracts. Leaves were extracted with ethanol and fractionated into n-hexane (Hex-F), ethyl acetate (Eth-F), and methanolic (Met-F) fractions using vacuum liquid chromatography. Phenolic, flavonoid, and saponin contents were quantified. Cytotoxicity was evaluated against four human cancer cell lines: A549 (lung), MCF-7 (breast), HeLa (cervical), and SKOV3 (ovarian), using the MTT assay. Apoptosis was assessed by Annexin V/PI flow cytometry, and genotoxicity or genoprotective effects were measured by the micronucleus assay in human lymphocytes. Phytochemical analysis revealed a total saponin yield of 29.95%. The Met-F fraction exhibited the highest concentrations of total phenolics (77.93 mg GAE/g) and triterpenoids (138.62 mg UAE/g), as well as the most pronounced cytotoxic activity, significantly reducing cell viability across all tested cancer lines in a concentration-dependent manner. The IC50 values were 108.23, 150.70, 213.77, and 781.07 µg/mL for SKOV3, MCF-7, HeLa, and A549 cell lines, respectively. In comparison, the Hex-F and Eth-F demonstrated weak cytotoxicity (IC50 > 1000 µg/mL). Flow cytometry suggested apoptosis as the predominant mode of cell death induced by Met-F. The micronucleus assay showed that Met-F reduced cisplatin-induced micronuclei formation in lymphocytes in a dose-dependent manner, indicating measurable genoprotective activity. These findings provide preliminary evidence that H. pastuchovii leaf fractions, particularly Met-F, possess in vitro cytotoxic and genoprotective properties that warrant further investigation.