Cynanchum acutum L. ethanolic fruit extract induces cell-cycle arrest and apoptosis in HepG2 human liver cancer cells via programmed cell death signaling pathways

Cynanchum acutum L. is a traditional medicinal plant known for its diverse pharmacological activities, such as anti-inflammatory, antibacterial, and antioxidant effects. This study aimed to evaluate the antiproliferative and apoptotic potential of the ethanolic extract of C. acutum L. fruits on liver cancer cells (HepG2), while assessing its safety in a normal human lung fibroblast cells (PCS-201-013). Comprehensive phytochemical profiling using HPLC-UV and LC-Ion trap-ESI-MS revealed, isovitexin (1407.5 mcg/g) and epicatechin (1175.3 mcg/g) as the dominant flavonoids, alongside substantial amounts of O-caffeic acid (170.4 mcg/g), orientin (162.6 mcg/g). In addition, vitexin (29.8 mcg/g), rutin (17.7 mcg/g), iso-orientin (21.2 mcg/g), and chlorogenic acid (15.6 mcg/g) were found in modest concentrations in dried fruit powder. The MTT assay exhibited a dose-dependent cytotoxicity with an IC₅₀ value of 40 μg/mL, while showing minimal toxicity toward normal fibroblast cells. It significantly suppressed cell migration and colony formation ability, indicating both anti-metastatic and antiproliferative effects. Moreover, the fruit extract resulted in mitochondrial membrane depolarization and increased ROS production in a dose-dependent manner. We also observed simultaneous cell cycle arrest at G1/S and G2/M phases and a significant increase in apoptotic cell populations (5.23 %–42.2 %). Collectively, these results highlight C. acutum fruit extract potential against the studied liver cancer cells. The presence of compounds, such as isovitexin and related flavonoids may have contributed to the observed effects. Future work will focus on isolating these compounds and conducting more detailed mechanistic and in vivo studies.