作者：胡建丽 ，章春芝，李倩，王晓芳，杜丽佳，王大伟

单位：河北北方学院附属第一医院 胸外科，河北 张家口 075000

Authors: Hu Jianli, Zhang Chunzhi, Li Qian, Wang Xiaofang, Du Lijia, Wang Dawei

Unit: Department of Thoracic Surgery, The First Affiliated Hospital of Hebei North University, Zhangjiakou 075000，Hebei, China

摘要：

目的 探讨黄芪皂苷Ⅳ（astragalosideⅣ，AS-Ⅳ）对食管癌TE-1细胞凋亡、干细胞样特性和PI3K/AKT通路的影响。方法 将TE-1细胞分为四组：空白对照组、黄芪皂苷Ⅳ20μg/ml组、黄芪皂苷Ⅳ40μg/ml组和黄芪皂苷Ⅳ80μg/ml组。给予各组细胞不同的黄芪皂苷Ⅳ药物浓度处理48 h后采用CCK-8法检测细胞存活率，成球实验检测干细胞成球情况，流式细胞术检测细胞凋亡，Western印迹法检测凋亡相关蛋白、干细胞标志蛋白和PI3K/AKT通路蛋白，以及免疫荧光检测AKT膜定位情况。结果 与空白对照组比较，黄芪皂苷Ⅳ40、80μg/ml组显著增加细胞凋亡率，显著上调caspase-3和caspase-9蛋白表达（P<0.05）；黄芪皂苷Ⅳ40、80μg/ml组显著减小干细胞成球球体直径及球体数量（P<0.05）；黄芪皂苷Ⅳ40、80μg/ml组显著下调干细胞标志蛋白SOX2、OCT4和CD44的表达（P<0.05）；黄芪皂苷Ⅳ40、80μg/ml组显著下调通路蛋白PI3K和AKT磷酸化水平，显著抑制AKT膜定位（P<0.05）。结论 黄芪皂苷Ⅳ可诱导TE-1细胞凋亡，抑制干细胞样特性，其作用机制可能与抑制PI3K/AKT通路活化有关。

关键词：黄芪皂苷Ⅳ；TE-1 细胞；细胞凋亡；干细胞特性；AKT 通路

Abstract：

Objective To investigate the effects of astragaloside Ⅳ（AS-Ⅳ） on apoptosis, stem cell-like characteristics and PI3K/AKT pathway of esophageal cancer TE-1 cells. Method TE-1 cells were divided into four groups: blank control group, astragaloside Ⅳ 20 μg/ml group, astragaloside Ⅳ 40 μg/ml group and astragaloside Ⅳ 80 μg/ml group. After 48 h of treatment with different concentrations of astragaloside Ⅳ, the cell survival rate was detected by CCK -8 method, the pellet formation of stem cells was detected by pellet formation assay, cell apoptosis was detected by flow cytometry, apoptosis -related proteins, stem cell marker proteins and PI3K/AKT pathway proteins were detected by Western Blot. And the localization of AKT membrane was detected by immunofluorescence. Result Compared with blank control group, astragaloside Ⅳ 40 and 80 μg/ml groups significantly increased the apoptosis rate, and significantly up-regulated the protein expression of caspase-3 and caspase-9 （P<0.05）. Astragaloside Ⅳ 40 and 80 μg/ml groups significantly decreased the diameter and number of stem cell spherules （P< 0. 05 ）; Astragaloside Ⅳ 40 and 80 μg/ml groups significantly down -regulated the expression of stem cell marker proteins SOX2, OCT4 and CD44 （P <0.05）. Astragaloside Ⅳ 40 and 80 μg/ml groups significantly down -regulated the phosphorylation levels of pathway protein PI3K and AKT, and significantly inhibited AKT membrane localization （P<0.05）. Conclusion Astragaloside Ⅳ can induce apoptosis of TE -1 cells and inhibit stem cell -like properties, which may be related to inhibition of PI3K/AKT pathway activation.

Key Words: Astragaloside Ⅳ; TE-1 cells; Apoptosis; Stem cell characteristics; AKT pathway