淫羊藿苷抑制PI3K/AKT通路对肺腺癌A549细胞存活和转移的影响  

Effect of Icariin on survival and metastasis of adenocarcinoma of lung cancer A549 cells via inhibiting PI3K/AKT pathway

作　　者：文艳梅[1] 梁宗安[2] 徐治波[1] 苟冶然 邓正旭[1] WEN Yan-Mei;LIANG Zong-An;XU Zhi-Bo;GOU Ye-Ran;DENG Zheng-Xu(Department of Respiratory Medicine,Chengdu Second People′s Hospital,Chengdu 610017,China)

机构地区：[1]成都市第二人民医院呼吸内科,成都610017 [2]四川大学华西医院呼吸与危重症医学科,成都610041

出　　处：《中国免疫学杂志》2020年第17期2091-2095,共5页Chinese Journal of Immunology

基　　金：四川省成都市卫生和计划生育委员会基金项目(2018032)。

摘　　要：目的:探讨淫羊藿苷对肺腺癌A549细胞存活和转移的影响及作用机制。方法:CCK8法检测不同浓度淫羊藿苷(0、0.1、0.25、0.5、1、2.5、5、10、20、50、100、200、300和400μmol/L)处理A549细胞后对其存活率的影响,并选择最佳浓度进行后续实验。流式细胞术检测细胞凋亡,划痕实验检测细胞迁移能力,Transwell检测细胞侵袭能力,Western blot法检测上皮标记蛋白E-cadherin和间质标记蛋白N-cadherin、Vimetin表达及PI3K和AKT的磷酸化情况;检测单独或联合给予10μmol/L PI3K激活剂740Y-P对细胞增殖、凋亡、侵袭、迁移和通路蛋白表达的影响。结果:100μmol/L及以上浓度的淫羊藿苷对A549细胞有明显的细胞毒性(P<0.05),选取10、20、50μmol/L作为后续实验浓度;淫羊藿苷可剂量依赖性提高A549细胞凋亡率(P<0.05),抑制细胞侵袭、迁移(P<0.05),影响N-cadherin、E-cadherin和Vimentin蛋白表达(P<0.05),下调PI3K和AKT磷酸化水平可减弱740Y-P对A549细胞增殖、侵袭迁移能力的增强作用,增强其对凋亡的抑制作用。结论:淫羊藿苷可通过影响PI3K/AKT信号通路激活抑制A549细胞存活和转移。Objective:To investigate effect of Icariin on survival and metastasis of human adencoarcinoma of lung cancer A549 cells.Methods:CCK8 assay was used to detect effects of different concentrations of Icariin(0,0.1,0.25,0.5,1,2.5,5,10,20,50,100,200,300 and 400μmol/L)on survival rate of A549 cells,and appropriate concentration was selected for subsequent experiments.Flow cytometry was used to detect cell apoptosis,scratch assay was used to detect cell migration.Transwell assay was used to detect cell invasion.Western blot was used to detect protein expressions of E-cadherin,N-cadherin,Vimetin and phosphorylation of PI3K and AKT.Effects of 10μmol/L PI3K activator 740Y-P alone or in combination on cell proliferation,apoptosis,invasion,migration and expression of pathway protein were examined.Results:Icariin(100μmol/L or above)had obvious cytotoxicity to A549 cells(P<0.05).Following experiments were conducted with 10,20 and 50μmol/L.Icariin could increase apoptotic rate of A549 cells in a dose-dependent manner(P<0.05),inhibit invasion and migration(P<0.05),and affect protein expressions of N-cadherin,E-cadherin and Vimentin(P<0.05).Down-regulating phosphorylation of PI3K and AKT were can weeken enhancement of proliferation,invasion and migration of A549 cells by 740Y-P,and increase inhibition of apoptosis.Conclusion:Icariin can inhibit survival and metastasis of A549 cells by activation of PI3K/AKT signaling pathway.

关 键 词：淫羊藿苷 人肺腺癌A549细胞 PI3K AKT 

分 类 号：R734.2[医药卫生—肿瘤]