柴胡皂苷A诱导Huh7细胞凋亡并上调细胞自噬水平的研究

Saikosaponin A induces apoptosis and upregulates autophagy in Huh7 cells

目的:探讨柴胡皂苷A(SSA)对人肝癌Huh7细胞凋亡和自噬水平的影响。方法:通过MTT和流式细胞术检测不同浓度SSA对Huh7细胞增殖和凋亡的影响;构建重组质粒pEGFP-N1-LC3B并转染Huh7细胞,SSA培养基干预后共聚焦显微镜下观察细胞自噬水平。Western Blot检测凋亡蛋白Bax、Bcl-2、PCNA及自噬相关蛋白LC3B、Beclin1、Apg12-Apg5的表达变化。结果:SSA可明显抑制Huh7细胞增殖,促进细胞凋亡,增加胞质内自噬小体形成数量,上调Bax、LC3B-Ⅱ、Beclin1、Apg12-Apg5的表达,下调Bcl-2、PCNA的表达。结论:SSA体外诱导Huh7细胞凋亡的同时上调了细胞自噬水平。

Objective:To study whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods:The effects of different concentrations of SSA on proliferation and apoptosis of Huh7 cells were detected by MTT and flow cytometry,and then constructed recombinant plasmid pEGFP-N1-LC3B and transfected into Huh7 cells.After intervened by SSA culture medium,the autophagy level was observed under confocal microscope.The expression of apoptosis proteins Bax,Bcl-2,PCNA and autophagy-related proteins LC3B,Beclin1,and Apg12-Apg5 were detected by Western Blot.Results:SSA can significantly inhibit the proliferation of Huh7 cells,promote apoptosis,increase the number of autophagy bodies in the cytoplasm,up-regulate the expression of Bax,LC3B-Ⅱ,Beclin1,Apg12-Apg5 and down-regulate the expression of Bcl-2,PCNA.Conclusion:SSA induced apoptosis of Huh7 cells in vitro and upregulated the autophagy level.