黄芪多糖通过诱导自噬作用促进结肠癌干细胞凋亡的机制  

Mechanism of astragalus polysaccharides promote colon cancer stem cell apoptosis by inducing autophagy

作　　者：李成军 沈光辉 LI Cheng-jun;SHEN Guang-hui(The Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学附属医院,咸阳712000

出　　处：《中华中医药杂志》2022年第4期2274-2279,共6页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:基于自噬作用探索黄芪多糖促进结肠癌干细胞凋亡及其相关分子机制。方法:免疫磁珠分选仪分选CD133、CD44共阳性结肠癌干细胞;悬浮培养验证CD133^(+)/CD44^(+)细胞干性;MTT检测黄芪多糖对CD133^(+)/CD44^(+)细胞增殖活性的影响;流式检测黄芪多糖对凋亡的影响;qPCR检测黄芪多糖对凋亡相关基因和自噬相关基因的影响;Western Blot检测黄芪多糖对凋亡相关蛋白和自噬相关蛋白的影响;免疫荧光检测各组细胞mTORC1蛋白的变化。结果:通过磁珠法成功富集CD133^(+)/CD44^(+)细胞,该细胞具有很强的成球能力。黄芪多糖可时间-浓度依赖地抑制CD133^(+)/CD44^(+)细胞增殖,并浓度依赖地诱导CD133^(+)/CD44^(+)细胞凋亡,促进Caspase 3、Caspase 9、Fas、Bax、ClassⅢPI3K、Beclin 1 mRNA和蛋白含量增加,降低Bcl-2、XIAP mRNA和蛋白表达。但是抑制自噬后,黄芪多糖对CD133^(+)/CD44^(+)细胞凋亡和自噬的诱导作用被逆转。结论:黄芪多糖可诱导结肠癌干细胞自噬,抑制其增殖,促进其凋亡。Objective:To explore the effects of astragalus polysaccharide in promoting colon cancer stem cell apoptosis and its related molecular mechanisms based on autophagy.Methods:Immunomagnetic bead sorter to sort CD133 and CD44co-positive colon cancer stem cells;suspension culture to verify the stemness of CD133^(+)/CD44^(+)cells;the effect of astragalus polysaccharide on the proliferation activity of CD133^(+)/CD44^(+)cells was detected by MTT;the effect of astragalu polysaccharide on apoptosis was detected by flow cytometry;the effect of astragalus polysaccharide on apoptosis-related and autophagyrelated genes was detected by qPCR;the effect of astragalus polysaccharide on apoptosis-related and autophagy-related proteins was edtected by Western Blot;the changes of mTORC1 protein in each group were detected by immunofluorescence.Results:CD133^(+)/CD44^(+)cells were successfully enriched by the magnetic bead method,and the cells have strong spheroidizing ability.Astragalus polysaccharides can inhibit the proliferation of CD133^(+)/CD44^(+)cells in a time-concentration-dependent manner,and induce apoptosis in CD133^(+)/CD44^(+)cells in a concentration-dependent manner,promote the mRNA and protein expression levels of Caspase 3,Caspase 9,Fas,Bax,ClassⅢPI3K and Beclin 1,and reduce the mRNA and protein expression levels of Bcl-2 and XIAP.But after inhibiting autophagy,the induction of Astragalus polysaccharide on CD133^(+)/CD44^(+)cell apoptosis and autophagy was reversed.Conclusion:Astragalus polysaccharide can induce autophagy of colon cancer stem cells,inhibit their proliferation,and promote their apoptosis.

关 键 词：黄芪多糖 自噬 结肠癌干细胞 凋亡 

分 类 号：R285[医药卫生—中药学]