三尖杉碱诱导肝癌细胞自噬的研究

Autophagy in liver carcinoma cell line HepG2 induced by cephalotaxine

目的探讨三尖杉碱抑制肝癌细胞增殖的方式及机制。方法采用MTT法检测三尖杉碱对肝癌细胞Hep G2增殖抑制作用;吖啶橙染色观察细胞形态;Westernblot检测自噬标志性蛋白及与自噬有关蛋白的表达。结果三尖杉碱以时间和剂量依赖方式抑制肝癌细胞Hep G2增殖;吖啶橙染色显示酸性囊泡细胞器增加;随着三尖杉碱作用浓度增加,自噬标志性蛋白LC3-Ⅱ以及Beclin-1表达上调。自噬抑制剂3-MA可抑制细胞增殖,下调蛋白LC3-Ⅱ以及Beclin-1表达。结论三尖杉碱通过上调Beclin-1诱导肝癌细胞发生自噬,从而抑制肝癌细胞生长。

Objective To investigate the way and the mechanism how cephalotaxine inhibits the proliferation of liver carcinoma cell line Hep G2. Cell viability of Hep MethodsG2 was detected using MTT assay. The morphology of cells was observed using acridine orange staining under a fluorescence microscope. The expression of proteins associated with autophagy was detected by Westernblot. Cephalotaxine inhibited the proliferation of Hep G2 in time-dependent and Resultsdose-dependent manner. Acridine orange staining showed the increase of cephalotaxine-induced acid autophagic vacuoles in Hep G2 cells. With the increase of cephalotaxine concentration, the expression of proteins LC3-Ⅱ and Beclin-1 was upregulated. Autophagy inhibitor 3-MA may inhibit the cell proliferation, and down-regulate the expression of LC3-Ⅱ and Beclin-1. Conclusion Cephalotaxine can induce autophagy in liver cancer cells by up-regulating Beclin-1 and thus inhibit the growth of liver cancer cells.