黄芪多糖诱导细胞自噬抑制食管癌EC109细胞增殖的研究  

Astragalus polysaccharide inhibits the proliferation of esophageal cancer EC109 cells by inducing cell autophagy

作　　者：常明智 张思雨 郝艳娇 张森露 张晓丽[1] CHANG Ming-zhi;ZHANG Si-yu;HAO Yan-jiao;ZHANG Sen-lu;ZHANG Xiao-li(Hebei North University,Zhangjiakou Hebei 075000;Sanquan College of Xinxiang Medical University,Xinxiang Henan 453003)

机构地区：[1]河北北方学院,河北张家口075000 [2]新乡医学院三全学院,河南新乡453003

出　　处：《中南药学》2022年第4期856-862,共7页Central South Pharmacy

基　　金：河北北方学院自然科学研究计划项目(No.YB2018008)。

摘　　要：目的研究黄芪多糖(APS)对食管癌EC109细胞增殖、凋亡、迁移和侵袭能力的影响及其与细胞自噬的关系。方法体外培养EC109细胞,实验分为以下几组:浓度为0(对照组)、1.0、1.5、2.0 mg·mL^(-1)的黄芪多糖组,5μmol·L^(-1) PIK-Ⅲ(自噬抑制剂)组,2.0 mg·mL^(-1)黄芪多糖联合5μmol·L^(-1) PIK-Ⅲ组。采用MTT法检测各组EC109细胞的增殖活性;吖啶橙染色实验和流式细胞术检测黄芪多糖组细胞的凋亡;扫描电镜观察黄芪多糖组EC109细胞表面结构的变化;Transwell细胞迁移、侵袭实验分别检测黄芪多糖组EC109细胞的迁移和侵袭能力;激光共聚焦显微镜检测黄芪多糖组LC3的荧光表达强度;Western blot检测黄芪多糖组自噬相关蛋白Beclin1、P62和LC3B的表达水平。结果与对照组相比,随着黄芪多糖浓度的升高,EC109细胞增殖抑制率及凋亡率显著增加;细胞表面结构破坏程度增加;细胞迁移和侵袭能力受到显著抑制;LC3荧光表达强度增强(P<0.01);P62蛋白表达水平显著降低(P<0.01),Beclin1和LC3B表达水平显著增高(P<0.05,P<0.01)。结论黄芪多糖可浓度依赖性地抑制EC109细胞的增殖、迁移和侵袭,并促进EC109细胞的凋亡,其机制可能与黄芪多糖诱导EC109细胞自噬有关。Objective To determine the effect of astragalus polysaccharide(APS)on the proliferation,apoptosis,migration and invasion of esophageal cancer EC109 cells and its relationship with cell autophagy.Methods EC109 cells were cultured in vitro and divided into 4 APS groups at the concentrations of 0(the control group),1.0,1.5 and 2.0 mg·mL^(-1),a 5μmol·L^(-1) PIK-Ⅲ(autophagy inhibitor)group and a 2.0 mg·mL^(-1) APS combined with 5μmol·L^(-1) PIK-Ⅲgroup.The proliferation of EC109 cells was determined by MTT assay.The degree of apoptosis of EC109 cells of the APS groups was determined by acridine orange staining assay and flow cytometry.Scanning electron microscope was used to observe the changes in the surface structure of EC109 cells of the APS groups.Transwell cell migration and invasion assay were used to detect the migration and invasion of EC109 cells in the APS groups,respectively.The fluorescence expression intensity of LC3 in the APS groups was detected by confocal laser microscopy.Western blot was used to detect the expression levels of the autophagy-related proteins Beclin1,P62 and LC3B in the APS groups.Results Compared with the control group,the proliferation inhibition rates and apoptosis rates of EC109 cells significantly increased with APS concentrations;the destruction of cell surface structure was increased;the migration and invasion were significantly inhibited (P < 0.01);and LC3 fluorescence expression intensity was enhanced (P < 0.01). The expression of P62 protein was significantly reduced (P<0.01), while the expression of Beclin1 and LC3B were increased (P<0.05, P < 0.01). Conclusion APS can inhibit the proliferation, migration and invasion of EC109 cells in a concentration-dependent manner and promote the apoptosis of EC109 cells, whose mechanism may be related to APS-induced autophagy in the EC109 cells.

关 键 词：黄芪多糖 食管癌EC109细胞 增殖 凋亡 自噬 

分 类 号：R285[医药卫生—中药学]