氧化苦参碱靶向miR-520e调控AEG-1介导的结肠癌细胞的转移、侵袭活性  

Effect of oxymatrine in targeting miR-520e to regulate the AEG-1-mediated metastasis and invasion of colon cancer cells

作　　者：易默[1] 吴友伟[1] 张健[1] 何瑜 刘倩[1] 史丽萍[1] 吕飒美[1] YI Mo;WU Youwei;ZHANG Jian;HE Yu;LIU Qian;SHI Liping;LV Samei(The Second Department of Gastroenterology,Shaanxi Provincial People’s Hospital,Xi’an 710068,China)

机构地区：[1]陕西省人民医院消化内二科,西安710068

出　　处：《吉林中医药》2022年第3期328-333,共6页Jilin Journal of Chinese Medicine

基　　金：陕西省自然科学基础研究计划项目(2018JM7118)。

摘　　要：目的探讨氧化苦参碱(OMT)通过微小RNA(miR)-520e调控星形胶质细胞瘤上调基因1(AEG-1)介导的结肠癌细胞的增殖、侵袭和迁移的影响。方法不同浓度OMT处理结肠癌细胞,MTT法检测细胞增殖抑制率;对数生长期的结肠癌细胞随机分为Control组、OMT组(含70μmol/L OMT的培养基培养24 h)、NC组(转染阴性对照片段24 h)、miR-520e inhibitor组(转染miR-520e抑制物24 h)和OMT+miR-520e inhibitor组(转染miR-520e抑制物24 h后,70μmol/L OMT处理24 h),MTT法检测细胞存活率,PCR检测miR-520e表达水平,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,双荧光素酶报告实验检测miR-520e与AEG-1靶向关系,蛋白印迹法检测AEG-1、白介素6(IL-6)和磷酸化信号转导子与激活子3(STAT3)蛋白相对表达量。结果随着OMT浓度的增加,结肠癌细胞增殖抑制率升高(P<0.05);与Control组比较,OMT组细胞存活率、侵袭细胞数以及AEG-1、IL-6和p-STAT3蛋白相对表达量降低,miR-520e表达水平以及划痕愈合率升高(P<0.05);与OMT组比较,OMT+miR-520e inhibitor组细胞存活率、侵袭细胞数以及AEG-1、IL-6和p-STAT3蛋白相对表达量升高,miR-520e表达水平以及划痕愈合率降低(P<0.05);与NC组比较,miR-520e inhibitor组侵袭细胞数以及AEG-1、IL-6和p-STAT3蛋白相对表达量升高,miR-520e表达水平以及划痕愈合率降低(P<0.05);与miR-520e inhibitor组比较,OMT+miR-520e inhibitor组细胞存活率、侵袭细胞数以及AEG-1、IL-6和p-STAT3蛋白相对表达量降低,miR-520e表达水平以及划痕愈合率升高(P<0.05)。结论OMT通过上调miR-520e抑制AEG-1表达,降低结肠癌细胞增殖、侵袭和迁移能力。Objective To investigate the eff ect of oxymatrine(OMT)on the proliferation,invasion and metastasis of colon cancer cells mediated by the astrocytoma elevated gene 1(AEG-1)via microRNA(miR)-520e.Methods Colon cancer cells were treated with diff erent concentrations of OMT,and the inhibition rate of cell proliferation was detected by MTT assay.Colon cancer cells in the logarithmic growth phase were randomly divided into a control group,an OMT group(cultured with 70μmol/L OMT for 24 h),an NC group(transfected with the negative control fragment for 24 h),an miR-520e inhibitor group(transfected with the miR-520e inhibitor for 24 h)and an OMT+miR-520e inhibitor group(transfected with the miR-520e inhibitor for 24 h,treated with 70μmol/L OMT for 24 h).MTT assay was used to detect the cell survival rate,PCR was used to detect the expression level of miR-520e,scratch assay to detect the cell metastasis ability,Transwell assay to detect the cell invasion ability,dual luciferase reporter assay to detect the targeting relationship between miR-520E and AEG-1,and Western blotting was used to detect the relative expression levels of AEG-1,interleukin 6(IL-6)and phosphorylated signal transductor and activator 3(STAT3)protein.Results With the increase of the OMT concentration,the proliferation inhibition rate of colon cancer cells were increased(P<0.05).Compared with the control group,the cell survival rate,the number of invasive cells and the relative expression levels of AEG-1,IL-6 and p-STAT3 protein were decreased in the OMT group,while the expression level of miR-520E and the scratch healing rate were increased(P<0.05).Compared with the OMT group,the cell survival rate,invasive cell number,relative expression levels of AEG-1,IL-6 and p-STAT3 protein were increased in the OMT+miR-520e inhibitor group,while the miR-520E expression level and scratch healing rate were decreased(P<0.05).Compared with the NC group,the number of invasive cells and the relative expression levels of AEG-1,IL-6 and p-STAT3 protein in the miR-520e i

关 键 词：结肠癌 氧化苦参碱 miR-520e AEG-1 

分 类 号：R735.3[医药卫生—肿瘤]