青蒿琥酯对肝癌细胞增殖的抑制作用和机制

Inhibitory effect and mechanism of artesunate on the proliferation of liver cancer cells

目的考察青蒿琥酯对肝癌细胞的增殖及其相关作用机制。方法应用CellTiter-Glo^■ Luminescent Cell Viability Assay活性试剂检测青蒿琥酯单独与索拉非尼联合用药对Huh7、HepG2细胞的活力。采用流式细胞术检测Huh7、HepG2细胞周期的变化,Western blotting检测AKT/mTOR/c-Myc信号通路相关蛋白的表达量。结果细胞活力检测结果显示Huh7、HepG2细胞的活力随作用时间-浓度增加而减少,在青蒿琥酯12.5μmol·L-1时,与索拉非尼联合应用,联合用药组对肝癌细胞生长的抑制率显著高于索拉非尼单药组,流式细胞的结果显示青蒿琥酯以浓度依赖的方式使Huh7、HepG2细胞的G2/M细胞周期比例逐渐增加,Western blotting结果表明青蒿琥酯单独作用肝癌细胞下调了p-AKT、p-S6、c-Myc蛋白水平。青蒿琥酯与索拉非尼联用显著地降低了肝癌细胞p-AKT、c-Myc水平。结论青蒿琥酯可能通过抑制AKT/mTOR/c-Myc信号通路使Huh7、HepG2细胞阻滞在G2/M期,并且青蒿琥酯可能通过降低p-AKT、c-Myc水平来增加了索拉非尼的敏感度。

Objective To investigate the inhibition of artesunate on the proliferation of hepatocellular carcinoma cells and its mechanism.Methods CellTiter-Glo^■Luminescent Cell Viability Assay was used to detect the activity of artesunate alone and in combination with sorafenib on Huh7 and HepG2 cells.Cell cycle changes were detected by flow cytometry,and the expression of AKT/c-Myc signaling pathway-related proteins was detected by Western blotting.Results The activity results showed that the viability of Huh7 and HepG2 cells decreased with the increase of the time-concentration of artesunate.12.5μmol·L-1 of artesunate was combined with sorafenib and the inhibitory effect was studied,the inhibitory rate was higher than that of the sorafenib group.The results of flow cytometry showed that artesunate gradually increased the G2/M cell cycle ratio of Huh7 and HepG2 cells in a concentration-dependent manner.Western blotting showed that p-AKT level,p-S6 level and c-Myc level decreased with the increase of the concentration when artesunate acts alone on hepatocellular carcinoma cells.It's found that a combination of artesunate and sorafenib reduced p-AKT level and c-Myc level in liver cancer cells.Conclusion Artesunate may block Huh7 and HepG2 cells in G2/M phase by inhibiting AKT/mTOR/c-Myc signaling pathway,and artesunate may increase the sensitivity of sorafenib by decreasing AKT phosphorylation level and c-Myc level.