黄芪多糖(Aps)与5-Fu联合对结肠癌细胞SW480凋亡诱导及分子机制研究  

The Apotosis Molecular Mechanism of SW480 Colon Cancer Cell Induced by the Combination of Astragalus Polysaccharide(Aps)and 5-Fu

作　　者：平娟[1] 周莉莉 李东换 王保全[1] 史朝辉[1] 李文倩 王熠宁 张永洲[1] PING Juan;ZHOU Li;LI Donghuan;WANG Baoquan;SHI Zhaohui;LI Wenqian;WANG Yining;ZHANG Yongzhou(The Huaihe Hospital of Henan University,Kaifeng,Henan 475004,China;The Kaifeng Children Hospital,Kaifeng,Henan 475004,China;The Yihe Hospital of Henan University,Zhengzhou,Henan 450009,China;Affiliated Cancer Hospital of Zhengzhou University,Zhengzhou,Henan 450009,China)

机构地区：[1]河南大学淮河医院,河南开封475004 [2]河南开封市儿童医院,河南开封475004 [3]河南大学附属郑州颐和医院,河南郑州450009 [4]郑州大学附属肿瘤医院,河南郑州450009

出　　处：《今日药学》2023年第9期652-656,664,共6页Pharmacy Today

基　　金：河南省科技厅项目(182102311244)。

摘　　要：目的初步探讨黄芪多糖(Aps)提高结肠癌SW480细胞对氟尿嘧啶(5-Fu)的敏感性,促进结肠癌细胞凋亡的分子机制。方法选择SW480结肠癌细胞株为实验对象,将黄芪多糖100μmol·L-1和5-Fu 30μg·mL-1组单用或联用48 h作为药物实验组,另设不加药为对照组,分别采用实时定量PCR和Western-bloting检测各组细胞bcl-2、p21、caspase-3、caspase-9基因表达和蛋白表达情况,比较阈值法确定基因的相对表达量,以GAPDH为内参,半定量分析蛋白相对表达量,EMSA法检测各组细胞核因子-κB的DNA结合活性,多组间样本均数比较采用单因素方差分析,组间样本均数比较采用LSD检验。结果联合用药组caspase-3、caspase-9和p21基因表达与蛋白表达均高于对照组、黄芪多糖组和5-Fu组,联合用药组生存素与蛋白表达均低于对照组、黄芪多糖组和5-Fu组,差异均具有统计学意义(P<0.05),caspase-3、p21在两单药组的表达与对照组相比,差异也具有统计学意义(LSD检验,P<0.05)。结论黄芪多糖与低剂量的5-Fu联用对结肠癌SW480细胞凋亡诱导作用显著大于高剂量5-Fu单独应用的效果,可能是通过黄芪多糖抑制核因子-κB的DNA结合活性,进一步上调caspase-3、caspase-9、p21及下调生存素起作用,提示黄芪多糖对5-Fu抗结肠癌细胞的作用具有协同和增强作用,为临床上结肠癌疾病的辅助化疗提供了一个新思路,黄芪多糖可为结肠癌化疗提供辅助治疗。OBJECTIVE To investigate the molecular mechanism of astragalus polysaccharide(Aps)to enhance the sensitivity of SW480 cells to fluorouracil(5-Fu)and promote the apoptosis of colon cancer cells.METHODS SW480 colon cancer cells were selected as the experimental subjects,and astragalus polysaccharide 100μmol·L^(-1)and 5-Fu 30μg·mL^(-1)groups were treated alone or in combination for 48 hours as the experimental group,and the other group was treated without drug addition as the control group.The Real-time quantitative PCR and western-bloting were used to detect the gene expression and protein expression of bcl-2,p21,caspase-3 and caspase-9 in each group,respectively.The relative expresion levels of genes were determined by comparative threshold method,the relative expression levels of proteins were semi-quantitatively analyzed with GAPDH as the internal reference,the DNA-binding activity of nuclear factor-κB in each group was detected by EMSA method.One-way analysis of variance was used to compare the sample means among multiple groups,LSD test was used for comparison of inter-group sample means.RESULTS The gene expression and protein expression of caspase-3,caspase-9 and p21 in combination treatment group were higher than those in control group,astragalus polysaccharide group and 5-Fu group.While the survivalin and protein expression in combination treatment group were lower than those in control group,astragalus polysaccharide group and 5-Fu group,with statistical significance(P<0.05).The expressions of caspase-3 and p21 in the two monotherapy groups were significantly different from those in the control group(LSD test,P<0.05).CONCLUSION The apoptosis induction effect of astragalus polysaccharide combined with low dose 5-Fu is significantly greater than that of higher dose 5-Fu along on colon cancer SW480 cells,which may be caused by inhibiting the DNA binding activity of nuclear factor-κB,further upregating caspase-3,caspase-9,p21 and down-regulating survival hormone.It is suggested that astragalus polysaccha

关 键 词：黄芪多糖 5-FU 结肠癌 细胞凋亡 分子机制 

分 类 号：R285[医药卫生—中药学]