黄芪多糖对肝癌HepG2细胞的抑制作用及其机制

The inhibition ability and its mechanism of polysaccharide of Astragalus in hepatic cancer HepG2 cell line

目的研究黄芪多糖(APS)和5-氟尿嘧啶(5-fu)联合使用对肝癌HepG2细胞的生长抑制及其作用机制。方法用RPMI1640培养液对低分化人肝癌细胞株HepG2进行传代培养,取对数生长期细胞,分为5组:A组:对照组,不含药物,使用同体积的磷酸盐缓冲液(PBS);B组:APS组,只给予100μg/mlAPS;C组:5-fu组,只给予100μg/ml5-fu;D组:5-fu+低浓度APS组,给予100μg/ml5-fu+100μg/mlAPS;E组:5-fu+高浓度APS组,给予100μg/ml5-fu+200μg/mlAPS。用噻唑蓝(MTT)法观察APS及5-fu对HepG2细胞增殖的影响;用流式细胞术方法测药物作用后细胞周期变化和细胞凋亡;western-blot检测凋亡相关蛋白表达。结果 APS和5-fu联合使用可抑制HepG2细胞的增殖,效应呈浓度和时间依赖性;APS和5-fu联合使用阻滞HepG2细胞周期于G1期;并可诱导HepG2细胞凋亡;APS和5-fu联合使用,可明显增高肝癌HepG2细胞中caspase-3、caspase-9蛋白的表达,同时检测到抑凋亡蛋白Bcl-2表达显著降低。在只使用5-fu的实验组细胞中均未检测到上述变化。结论 APS和5-fu联合使用能抑制肝癌HepG2细胞株的增殖,其机制之一是影响细胞周期使之阻滞于G1期,激活了细胞凋亡系统,从而诱导细胞凋亡。

Objective To observe the effect of cell inhibiting and promoting apoptosis of polysaccharide of Astragalus（APS）on hepatic cancer HepG2 cell line.Methods HepG2 cells were maitained in RPMI1640 culture solution.After culture logarithmic phase cells were divided into 5 groups and treated with different pharmaceuticals：A group ：control（Cont）;B group ：100 μg/ml APS;C group ：100 μg/ml 5-fu;D group ：100 μg/ml 5-fu＋100 μg/ml APS;E group ：100 μg/ml 5-fu＋200 μg/ml APS.cell proliferation effection were detected by MTY assay ;the cell cycle and apoptosis were studied by flow cytometry（FCM）;apoptosis protein expression were examined by western blot,respectively.Results Astragalus polysaccharides combined with 5-fu could inhibit cell proliferation of HepG2 cells.The inhibitory effect showed that a dose 0-time-dependent manner.Astragalus polysaccharides could retard cell cycle at G1 stage;Astragalus polysaccharides could induce apoptosis of HepG2 cells,could increase the expression of caspase-3,caspase-9,and decrease the expression of Bcl-2 in HepG2 cells.Conclusions Astragalus polysaccharides combined with 5-fu could inhibit cell proliferation of HepG2 cells,which might be due to the retardarce of cell cycle at G1 stage.