摘要
目的探究赤芍总苷(total paeony glycoside,TPG)对人肺癌细胞增殖、迁移和侵袭的影响,并进一步研究其潜在的调控机理。方法以人肺癌A549细胞作为细胞研究模型,分别给予0、50、100、200μg·mL^-1的TPG处理12、24 h,进行时间-剂量效应分析,并选定100μg·mL^-1TPG处理24 h为机制研究模型;采用小干扰RNA(si AKT)干预Akt蛋白表达水平,噻唑蓝(MTT)法检测不同浓度和时间点TPG对人肺癌A549细胞增殖抑制的影响;采用蛋白免疫印迹法(Western Blot)分别检测Akt、MMP2、p-Akt和MMP9蛋白表达的水平;采用细胞划痕检测TPG对人肺癌A549细胞迁移的能力;采用Transwell实验检测TPG对A549细胞迁侵袭能力的影响。结果MTT试验显示:TPG对A549细胞的生长呈剂量和时间依赖性抑制作用;Western Blot结果显示:与对照组比较,各处理组随着TPG剂量递增,Akt、p-Akt、MMP2和MMP9蛋白表达的水平呈现剂量依赖性下降;细胞划痕结果显示:TPG可呈剂量依赖性抑制A549细胞的迁移;Transwell试验表明:TPG可抑制A549细胞的侵袭。采用si AKT联合TPG处理,细胞划痕和Transwell试验发现:与TPG单独处理组比较,TPG+si AKT联合处理抑制A549细胞的迁移和侵袭更显著。Western Blot检测发现:与TPG单独处理组比较,TPG+si AKT联合处理组Akt、p-Akt、MMP2和MMP9蛋白表达的水平均显著下降。结论TPG可抑制人肺癌A549细胞的增殖、迁移和侵袭,其分子机制可能是通过抑制Akt通路的激活,进而降低MMP-2和MMP-9蛋白的表达,研究结果为TPG的临床应用提供了实验依据。
OBJECTIVE To investigate the effect of total paeony glycoside(TPG)on proliferation,migration and invasion of A549 human lung cancer cells,and to explore its potential molecular mechanism of action.METHODS A549 human lung cancer cells were treated with TPG(0,50,100,200μg·mL^-1)for 12 h and 24 h respectively for dose and time dependent effect analysis,100μg·mL^-1TPG treatment for 24 h was selected as the mechanism study model,and small interfering RNA(siAKT)was used to intervene the expression of Akt protein,and thiazolyl blue(MTT)method was used to detect the effect of TPG on the proliferation of A549 cells at different concentrations and time points.Western Blotting was used to detect Akt,p-Akt,MMP2 and MMP9 protein expression levels.Cell scratches and transwell assays were used to detect the effect of TPG on migration and invasion of lung cancer cells.RESULTS MTT assay results showed that TPG inhibited the growth of A549 cells in a dose and time dependent manner.Western Blot results showed that,compared with Ctrl group,the protein expression levels of Akt,p-Akt,MMP2 and MMP9 were significantly decreased by a dose dependent manner in TPG group.Cell scratches and Transwell assay results showed that TPG inhibited the migration and invasion of A549 cells in a dose dependent manner.Using siAKT combined with TPG treatment,cell scratches and Transwell assay,it has found that TPG+siAKT combined treatment group inhibited the migration and invasion of A549 cells more significantly than TPG alone treatment group.Western Blot analysis showed that the expression of Akt,p-Akt,MMP2 and MMP9 in TPG+siAKT treatment group was significantly lower than that in TPG alone treatment group.CONCLUSION TPG inhibits proliferation,migration and invasion of A549 human lung cancer cells by the Akt pathway,providing a theoretical basis for the clinical treatment of lung cancer.
作者
高晓会
张亚利
张治业
郭双双
郭艳珍
陈小兵
GAO Xiaohui;ZHANG Yali;ZHANG Zhiye;GUO Shuangshuang;GUO Yanzhen;CHEN Xiaobin(Department of Medical Oncology,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang,Henan,471003 P.R.China)
出处
《华西药学杂志》
CAS
CSCD
2019年第3期243-248,共6页
West China Journal of Pharmaceutical Sciences
基金
国家自然科学基金资助项目(批准号:81172240)
