翼核果素对肝癌HepG2细胞凋亡及细胞骨架蛋白F-actin的影响

Effects of Ventilagolin On the Apoptosis of Human Hepatoma Hep G2 Cells and Cytoskeletal Protein F-actin

目的:研究翼核果素对人肝癌Hep G2细胞增殖和凋亡的影响,并探讨其可能的机制。方法:以不同浓度的翼核果素作用于体外培养的Hep G2细胞,采用MTT法检测细胞生长抑制率,HE、吖啶橙染色观察细胞形态,流式细胞仪检测细胞凋亡率和周期,激光共聚焦显微镜观察细胞骨架蛋白F-actin形态学变化。结果:翼核果素可明显抑制人肝癌Hep G2细胞的增殖且呈剂量依赖性,作用于Hep G2细胞24 h的IC50值为124.77μmol·L^(-1)。显微镜可见细胞核变形、染色体聚集等典型的凋亡形态学改变。流式细胞仪检测结果显示,60、90μmol·L^(-1)翼核果素给药组细胞凋亡的百分率显著高于对照组,G2/M期细胞比例显著高于对照组,细胞骨架蛋白F-actin呈现解聚断裂状态。结论:翼核果素可抑制Hep G2细胞的增殖,诱导细胞凋亡,其机制可能与细胞周期阻滞、细胞骨架蛋白F-actin解聚有关。

Objective： To study the inhibition effects of ventilagolin and proliferation and apoptosis on human hepatocellular carcinoma cells HepG2, discusses the possible mechanisms. Methods： HepG2 cells in culture medium were treated with different concentrations of ventilagolin. The inhibitory rate of the ceils was measured by MTT assay. The morphology of cells was observed by HE and Acridine orange stain. Cell cycle changes and apoptotic rate were detected by Flow Cytometry （FCM）. The mophological changes of cytoskeletal protein F-actin were observed by laser confocal microscopy. Results： The growth of HepG2 cells was significantly inhibited by ventilagolin, and the growth inhibitory rate increased with increasing concentration. IC50 of ventilagolin was 124. 77 μmol. L^- 1 after 24 hours. A characteristic apoptosis change including nuclear disintergration and chromatin agglomerate was displayed. The apoptosis rate of HepG2 cell treated with 60, 90 μmol· L^-1 ventilagolin was significantly higher than that of the control group, and the percentage of G2/M phase of I-IepG2 cell was also significantly higher than that of the control group. F-actin eytoskeletal protein was showed from polymerization to depolymerization after ventilagolin treatment. Conclusion： Ventilagolin can obviously inhibit the proliferation and induce apoptosis of HepG2 cells. The mechanisms may be due to blocking cell cycle and depolymerizating cytoskeletal protein F-actin.