黄芪多糖对阿帕替尼抗肝癌细胞的增强作用及对CA199、CA724肿瘤标志物的影响

Enhancement of Astragalus Polysaccharide on Apatinib Against Liver Cancer Cells and Its Effect on Tumor Markers of CA199 and CA724

目的 探讨黄芪多糖对阿帕替尼抗肝癌活性以及CA199、CA724肿瘤标志物的影响。方法 采用CCK-8法检测黄芪多糖(50、100、200、400 mg/L)、阿帕替尼(10、20、40、80μmol/L)以及黄芪多糖联合阿帕替尼[3个浓度(100、200、400)黄芪多糖+20μmol/L阿帕替尼]对肝癌细胞存活率的影响。根据细胞存活率将肝癌细胞Hep3B分为对照组、200 mg/L黄芪多糖组、20μmol/L阿帕替尼组、200 mg/L黄芪多糖+20μmol/L阿帕替尼组。采用流式细胞术、Transwell法检测Hep3B凋亡率以及迁移、侵袭细胞数。ELISA法检测细胞培养液中糖类抗原199(CA199)、CA724水平。结果 对照组、200 mg/L黄芪多糖组、20μmol/L阿帕替尼组、200 mg/L黄芪多糖+20μmol/L阿帕替尼组Hep3B细胞凋亡率分别为(5.12±0.26)%、(8.92±0.47)%、(13.96±0.75)%、(37.73±1.95)%;细胞迁移数量分别为(274±12.23)个、(220±13.50)个、(198±10.83)个、(114±5.54)个;细胞侵袭数量分别为(188±7.65)个、(152±10.19)个、(127±8.53)个、(82±3.047)个;CA199水平分别为(28.43±1.67)U/ml、(23.56±2.16)U/ml、(20.29±1.25)U/ml、(5.42±0.22)U/ml,CA724水平分别为(68.96±3.49)U/ml、(60.45±3.26)U/ml、(56.97±1.70)U/ml、(26.16±1.65)U/ml。与对照组比较,200 mg/L黄芪多糖组、20μmol/L阿帕替尼组细胞存活率、迁移和侵袭数、CA199和CA724水平降低(P<0.05),凋亡率升高(P<0.05);与200 mg/L黄芪多糖组、20μmol/L阿帕替尼组比较,200 mg/L黄芪多糖+20μmol/L阿帕替尼组细胞存活率、迁移和侵袭数、CA199和CA724水平降低(P<0.05),凋亡率升高(P<0.05)。结论 黄芪多糖可增强阿帕替尼对肝癌细胞的抗增殖、促凋亡和抗迁移侵袭作用,并降低肿瘤标志物CA199、CA724的水平。

Objective To investigate the effect of astragalus polysaccharide on the anti-hepatocellular carcinoma activity of apatinib and the tumor markers of CA199 and CA724.Methods The effect of Astragalus polysaccharides(50,100,200,400 mg/L),apatinib(10,20,40,80μmol/L)and astragalus polysaccharides combined with apatinib[3 concentrations(100,200,400 astragalus polysaccharide+20μmol/L apatinib)]on the survival rate of liver cancer cells were detected by CCK-8 method.According to the cell survival rate,the liver cancer cells Hep3B were divided into control group,200 mg/L astragalus polysaccharide group,20μmol/L apatinib group,200 mg/L astragalus polysaccharide+20μmol/L apatinib group.Flow cytometry and Transwell method were used to detect the apoptosis rate of Hep3B and the number of migrating and invasive cells.The levels of carbohydrate antigen 199(CA199)and CA724 in cell culture medium were detected by ELISA.Results The apoptosis rate of Hep3B cells in the control group,200 mg/L astragalus polysaccharide group,20μmol/L apatinib group,200 mg/L astragalus polysaccharide+20μmol/L apatinib group Hep3B were(5.12±0.26)%,(8.92±0.47)%,(13.96±0.75)%,(37.73±1.95)%;the number of cell migration were(274±12.23),(220±13.50),(198±10.83),(114±5.54);the number of cell invasion were(188±7.65),(152±10.19),(127±8.53),(82±3.047);CA199 level was(28.43±1.67),(23.56±2.16),(20.29±1.25),(5.42±0.22)U/ml,CA724 levels were(68.96±3.49),(60.45±3.26),(56.97±1.70),(26.16±1.65)U/ml.Compared with the control group,the cell survival rate,migration and invasion numbers,CA199 and CA724 levels in the 200 mg/L astragalus polysaccharide group and 20μmol/L apatinib group were decreased(P<0.05),cell apoptosis rate was increased(P<0.05).Compared with the 200 mg/L astragalus polysaccharide group and the 20μmol/L apatinib group,the cell viability,migration and invasion numbers,CA199 and CA724 levels in the 200 mg/L astragalus polysaccharide+20μmol/L apatinib group were decreased(P<0.05),and the apoptosis rate was increased(P<0.05).Conclusions Astragalus polysaccharide could enhance the anti-proliferation,pro-apoptosis and anti-migration and invasion of apatinib on liver cancer cells,and reduce the levels of tumor markers CA199,CA724.