拓扑异构酶Ⅱ抑制剂GL331对HepG2细胞体外生长的抑制作用

GL331,a topoisomerase Ⅱ inhibitor,induces growth arrest in HepG2 cells in vitro

目的:研究拓扑异构酶Ⅱ抑制剂GL331体外对人肝癌HepG2细胞的生长抑制作用及其机制。方法:采用MTT法测定GL331对人肝癌HepG2细胞存活率的影响;克隆原形成法检测对细胞集落形成的影响;DAPI染色观察细胞形态学变化;流式细胞术检测细胞凋亡百分率的变化;蛋白免疫印迹技术观察Bax,Bcl-2,Fas,FasL,Akt/p-Akt,caspase-3,clea-caspase等相关蛋白水平变化;caspase-3酶活性检测试剂盒检测对蛋白酶caspase-3活性的影响。结果:GL331可以浓度和时间依赖性地抑制HepG2细胞的增殖,同时促进HepG2细胞凋亡,提高Bax/Bcl-2比率,增加FasL的表达,降低p-Akt的表达。结论:GL331对HepG2细胞的增殖有明显的抑制作用,其作用机制与诱导HepG2细胞发生凋亡,调节凋亡相关蛋白的表达,并抑制存活通路相关蛋白的活化有关。

Objective：To observe the effects of GL331,a topoisomerase Ⅱ inhibitor,on the growth of HepG2 cells.Methods：MTT and Colony-forming assay were carried out to explore the effects of GL331 on the cellular proliferation and the colony formation.Morphological changes were observed by DAPI staining.The apoptosis was then analyzed by Flow Cytometry.Apoptosis-and survival-related molecules were observed by western blotting assay.Caspase-3 activity was measured by Caspase-3 activity assay.Results：GL331 suppressed the proliferation of HepG2 cells in a concentration-and time-dependent manner.GL331 induced apoptosis through down-regulating apoptotic protein Bcl-2,up-regulating the Bax/Bcl-2 ratio and pro-apoptotic protein FasL,and finally activating the Caspase-3 enzyme.Meanwhile,GL331 could decrease the level of p-Akt.Conclusion：GL331 arrests the growth of HepG2 cells in vitro,and the mechanisms may be associated with initiating apoptosis-related pathways and inhibiting the survival-related protein.