The anti-angiogenic and cytotoxic potential of different Eremurus spectabilis fractions on hepatocellular carcinoma (Hep3B) cells: An in vitro, in Ovo, and in silico study

Background: Hepatocellular carcinoma (HCC) remains a major health concern, with angiogenesis playing a key role in its progression. Medicinal plants offer valuable anticancer potential. Eremurus spectabilis (ES), traditionally used in folk medicine, has not been fully explored for its anticancer properties. This study investigates its cytotoxicity on Hep3B cells and its anti-angiogenic activity. Methods: E. spectabilis fractions (hexane, chloroform, ethyl acetate, and aqueous ethanol) were obtained from the ethanolic extract. The total phenolic content (TPC) was measured, and the active fractions were analyzed using gas chromatography-mass spectrometry (GC-MS). The xCELLigence Real-Time Cell Analyzer was used to evaluate cytotoxicity against Hep3B cells. Anti-angiogenic activity was assessed using the CAM assay, and docking studies were conducted to predict the mechanism of anti-angiogenesis. Results: ESEA (154.80 ± 0.10 mg/g) had the highest phenolic content, followed by ESC (84.81 ± 6.81 mg/g). Palmitic acid (26%), khusimyl acid (15.9%), glycerol (12.1%), and D-(+)-talofuranose pentakis(trimethylsilyl) ether (9.7%) were the major compounds in ESC, while D-glucopyranose (19.4%), β-D-(+)-talopyranose (15.7%), and D-(−)-tagatofuranose pentakis(trimethylsilyl) ether (13.6%) were among the major compounds in ESEA. All fractions revealed significant cytotoxicity on Hep3B cells, with ESEA (24-h IC₅₀: 6.53 μg/mL) and ESAE (24-h IC₅₀: 7.89 μg/mL) being more effective. ESC also exhibited strong anti-angiogenic effects by reducing vessel area, length, and branching points. These findings were supported by molecular docking. The major phytochemicals significantly interact with VEGFR-2 and FGFR1. Conclusions: E. spectabilis shows a promising dual action for cancer therapy. However, further research is necessary to predict the exact mechanism of action.