黄芪多糖通过线粒体自噬抑制肝癌肿瘤细胞  

Astragalus polysaccharides inhibit hepatocellular carcinoma cells through mitochondrial autophagy

作　　者：张慧蓉 钱敏[2] 李萌 ZHANG Huirong;QIAN Min;LI Meng(Department of Clinic Laboratory,Zibo Bashan Wanjie Hospital,Zibo 255200,China;Department of Clinic Laboratory,Jinan First People′s Hospital,Jinan 250031,China;Department of Clinic Laboratory,the Third Hospital of Shandong Province,Jinan 250031,China)

机构地区：[1]淄博岜山万杰医院检验科,淄博255200 [2]济南市第一人民医院检验科,济南250031 [3]山东省立第三医院检验科,济南250031

出　　处：《西北药学杂志》2021年第3期426-429,共4页Northwest Pharmaceutical Journal

基　　金：淄博市基因组编辑与细胞应用研究平台开放课题(编号:2019GECA006)。

摘　　要：目的研究黄芪多糖(APS)对肝癌细胞的抑制作用及其抑制作用是否涉及自噬。方法应用克隆形成法和溴化噻唑蓝四氮唑(MTT)法检测APS对肝癌细胞SMMC-7721的生长抑制作用,用流式细胞术检测APS对SMMC-7721细胞周期的影响,Western Blot法检测APS对自噬相关蛋白的表达。结果质量浓度为0.75、1.50、3.00 mg·mL^(-1)的APS均可减少SMMC-7721的细胞克隆数,且随质量浓度的增加抑制作用明显增强;APS质量浓度梯度地抑制SMMC-7721细胞的存活率,且随着时间增加APS对SMMC-7721细胞的抑制作用也增强。APS产生作用后,S期细胞比例减少,而G0/G1期细胞比例增加。此外,3.00 mg·mL^(-1)的APS可显著增加微管相关轻链3B(LC3B)、Beclin 1和p62等自噬相关蛋白的表达,提示APS可刺激自噬;当自噬抑制剂3-甲基腺嘌呤(3-MA)和巴佛洛霉素A1(Baf)存在时,APS抑制SMMC-7721细胞作用消失。结论APS对肝癌细胞SMMC-7721有显著的抑制作用,诱导细胞阻滞于G0/G1期可能是其机制之一,且其抑制作用可由自噬介导。Objective To investigate the inhibitory effect and mechanism of astragalus polysaccharides(APS)on hepatocellular carcinoma,and to investigate whether its inhibitory effects involve autophagy.Methods Clonogenic assay and tetrazolium bromide(MTT)were used to detect the inhibitory effect of APS on the growth of SMMC-7721 cells.Flow cytometry was used to detect the cell cycle of SMMC-7721 cells.Western Blot was performed to evaluate the expression of autophagy-related proteins.Results APS of 0.75,1.50,3.00 mg·mL^(-1) could reduce the cell clone number of SMMC-7721,and the inhibitory effect increased with the increase of concentration;APS dose-dependently inhibited the survival rate of SMMC-7721 cells;the inhibitory effect of APS on SMMC-7721 cells increased with time.After APS treatment,the proportion of cells in S phase decreased,while that in G0/G1 phase increased.In addition,APS at 3.00 mg·mL^(-1) significantly increased the expression of autophagy related proteins including microtube associated light chain 3B(LC3B),Beclin 1 and p62,suggesting that APS could stimulate autophagy.It should be noted that APS could inhibit SMMC-7721 cells in the presence of autophagy inhibitors 3-methyladenine(3-MA)and bafilomycin A1(Baf).Conclusion APS inhibit the growth of SMMC-7721 cells,through inducing the cell arrest at G0/G1 phase,and its inhibitory effect may be mediated by autophagy.

关 键 词：黄芪多糖(APS) SMMC-7721细胞 自噬 增殖 凋亡 

分 类 号：R965[医药卫生—药理学]