Abstract: In order to study the effect of citrusinol on human hepatocellular carcinoma cells HepG₂,terazolium salt colorimetry assay(MTT ),cell growth curve and colony formation assay was used to detected the proliferation and colony formation.The morphology of cells was observed by HE staining and Acridine orange staining.Cell cycle changes were detected by Flow Cytometry(FCM).The morphological changes of cytoskeletal protein F-actin were observed by laser confocal microscopy.The results showed thatcitrusinol inhibited HepG₂cells proliferation and colony formation in a dose-dependent manner.IC₅₀ of citrusinol was 79.14 μmol/L after 24 hours.The morphology of HepG₂ cells was changed,showing cell shrinkage,cytoplasm vacuoles and nuclear chromatin pyknosis.The percentage of G2/M phase of HepG₂ cells treated with 70,80 μmol/L citrusinol was significantly higher than that of the control group (22.72±1.07,28.68±1.36 vs 16.34±0.66,P<0.01).F-actin cytoskeletal protein was showed from polymerization to depolymerization after citrusinol treatment.Based on the experimental results,it was concluded that citrusinol can prevent cell mitosis to arrest the cell cycle in G2/M phase,by disrupting Actin cytoskeleton,which may be one of the mechanisms of citrusinol's anti-tumor effect.

Key words: citrusinol, HepG₂ cell, antiproliferation, cell cycle, cytoskeletal protein