黄芪皂苷Ⅱ联合5-氟尿嘧啶处理抑制肝癌细胞增殖的实验研究

Mechanism of Astragaloside Ⅱ combined with 5-fluorouracil in inhibiting liver cell proliferation

目的:研究黄芪皂苷Ⅱ联合5-氟尿嘧啶处理对肝癌细胞增殖的影响并分析具体的分子机制。方法:培养肝癌细胞株HepG2,取对数期生长的细胞并用不同条件进行处理,空白对照组用不含血清的DMEM进行处理,5-Fu处理组用0.5、1、5、10μmol/L 5-Fu处理,5-Fu联合黄芪皂苷II处理组用10μmol/L 5-Fu联合20、40、80μmol/L黄芪皂苷II处理。处理后24、48、72h时,采用MTS试剂盒测定细胞活力;处理后24h时,采用Elisa试剂盒测定细胞内凋亡蛋白的含量。结果:与空白对照组比较,5-Fu处理能够以时间依赖性和剂量依赖性的方式降低肝癌细胞存活率;与10μmol/L 5-Fu单药处理组比较,10μmol/L 5-Fu联合黄芪皂苷II处理能够以时间依赖性和剂量依赖性的方式降低肝癌细胞存活率;10μmol/L 5-Fu处理后的Fas、FasL、Bax、Caspase-3、Caspase-8、Caspase-9表达量显著高于空白对照组,10μmol/L 5-Fu联合40μmol/L黄芪皂苷II处理后的Fas、FasL、Bax、Caspase-3、Caspase-8、Caspase-9表达量显著高于10μmol/L 5-Fu处理组。结论:黄芪皂苷II联合5-氟尿嘧啶处理能够增强5-氟尿嘧啶对肝癌细胞增殖的抑制效果,介导该作用的凋亡蛋白是Fas/FasL及Bax。

[ABSTRACT]Objective：To study the mechanism of Astragaloside II combined with 5-fluorouracil in inhibiting liver cell proliferation and to analyze the specific molecular mechanisms.Methods：Liver cancer cell lines HepG2 were cultured,cells of logphase growth were collected and treated with different conditions,blank control group were treated with DMEM without se-rum,5-Fu treatment group were treated with 0.5,1,5 and 10 μmol/L 5-Fu,and 5-Fu combined with Astragaloside II treat-ment group were treated with 10 μmol/L 5-Fu combined with 20,40 and 80 μmol/L Astragaloside II.24,48 and 72h after treatment,MTS kit was used to determine the cell viability;24h after treatment,Elisa kit was used to determine the content of apoptosis protein in cells.Results：Compared with blank control group,5-Fu treatment could reduce the survival rate of livernbsp;cancer cells in time-dependent and dose-dependent manner;compared with 10 μmol/L 5-Fu single drug treatment group,10μmol/L 5-Fu combined with Astragaloside II treatment could reduce the survival rate of liver cancer cells in time-dependent and dose-dependent manner;Fas,FasL,Bax,Caspase-3,Caspase-8 and Caspase-9 expression levels after 10 μmol/L 5-Fu treat-ment were significantly higher than those of blank control group,and Fas,FasL,Bax,Caspase-3,Caspase-8 and Caspase-9 expression levels after 10 μmol/L 5-Fu combined with 40 μmol/L Astragaloside II treatment were significantly higher than those of 10μmol/L 5-Fu treatment group.Conclusion：Astragaloside II combined with 5-fluorouracil treatment can enhance the inhibitory effect of 5-fluorouracil on liver cancer cell proliferation through mediating apoptosis proteins of Fas/FasL and Bax.