槐定碱对肺腺癌细胞A549顺铂敏感性及miR-216a-5p/ZEB1轴的影响  

Effect of Sophoridine on Cisplatin Sensitivity and miR-216a-5p/ZEB1 Axis in Lung Adenocarcinoma Cell A549

作　　者：聂佳 李方 李延波 NIE Jia;LI Fang;LI Yan-Bo(Dept.of Respiratory Medicine,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050000 Hebei,China;Dept.of Oncology,Handan Central Hospital,Handan 056000 Hebei,China;Dept.of General Internal Medicine,Xindu District People’s Hospital,Xingtai 054001 Hebei,China)

机构地区：[1]河北省中医院呼吸内科,河北石家庄050000 [2]邯郸市中心医院肿瘤科,河北邯郸056000 [3]邢台市信都区人民医院普通内科,河北邢台054001

出　　处：《广州中医药大学学报》2022年第7期1650-1657,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：河北省医学科学研究课题计划项目(编号:20200465)。

摘　　要：【目的】观察槐定碱对肺腺癌A549细胞耐药性和敏感性的影响。【方法】(1)将肺腺癌A549细胞随机分为对照组、槐定碱组、顺铂组和槐定碱+顺铂组。对照组常规培养,槐定碱组和顺铂组分别加入40μmol/L的槐定碱和2.0 nmol/L的顺铂,槐定碱+顺铂组同时加入40μmol/L槐定碱和2.0 nmol/L顺铂,处理24 h后,平板克隆实验测定细胞集落形成能力,流式细胞仪测定细胞凋亡率。(2)将肺腺癌A549细胞随机分为对照(control)组,抑制剂阴性对照(inhibitor-NC)组、抑制剂(inhibitor)组、槐定碱+inhibitor-NC组、槐定碱+inhibitor组、槐定碱组,相应干预后,实时定量聚合酶链反应(RT-qPCR)法检测细胞中miR-216a-5p和锌指E盒结合蛋白1(ZEB1)mRNA表达,双荧光素酶报告基因系统验证miR-216a-5p与ZEB1的靶向关系,Western Blot法检测细胞中ZEB1、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达。【结果】(1)与对照组比较,槐定碱组、顺铂组和槐定碱+顺铂组细胞集落数均减少,细胞凋亡率升高(均P<0.05);槐定碱+顺铂组细胞集落数少于槐定碱组和顺铂组,细胞凋亡率高于槐定碱组和顺铂组(均P<0.05)。(2)与control组和inhibitor-NC组比较,inhibitor组miR-216a-5p表达及Bax蛋白表达降低,ZEB1 mRNA和蛋白及Bcl-2蛋白表达水平升高(均P<0.05);与control组比较,槐定碱组miR-216a-5p表达及Bax蛋白表达水平升高,ZEB1 mRNA和蛋白及Bcl-2蛋白表达水平降低(均P<0.05);与inhibitor组比较,槐定碱+inhibitor组miR-216a-5p表达及Bax蛋白表达水平升高,ZEB1 mRNA和蛋白及Bcl-2蛋白表达水平降低(均P<0.05);与槐定碱组、槐定碱+inhibitor-NC组比较,槐定碱+inhibitor组miR-216a-5p表达及Bax蛋白表达水平降低,ZEB1 mRNA和蛋白及Bcl-2蛋白表达水平升高(均P<0.05)。双荧光素酶实验结果显示,miR-216a-5p可靶向调控ZEB1表达。【结论】槐定碱可增强A549细胞对顺铂的敏感性,亦可通过上调miR-216a-5p靶向降低ZEB1表达,发挥抗肺Objective To observe the effect of sophoridine on drug resistance and sensitivity of lung adenocarcinoma A549 cells.Methods(1)The A549 cells were randomly divided into the control group,sophoridine group,cisplatin group and sophoridine+cisplatin group.Routine culture was performed in the control group,40μmol/L of sophoridine and 2.0 nmol/L of cisplatin were added to the sophoridine group and cisplatin group respectively,and both 40μmol/L of sophoridine and 2.0 nmol/L of cisplatin were added simultaneously to the sophoridine+cisplatin group.After 24 hours,colony formation ability was detected by plate cloning assay,and apoptosis rate was detected by flow cytometry.(2)A549 cells were randomly divided into control group,inhibitor-NC group,inhibitor group,sophoridine+inhibitor-NC group,sophoridine+inhibitor group and sophoridine group.After the corresponding intervention,the expression of miR-216a-5p and Zinc finger E-box binding protein 1(ZEB1)mRNA in A549 cells was detected by real-time quantitative polymerase chain reaction(RTqPCR).Dual luciferase reporter gene system was used to verify the targeting relationship between miR-216a-5p and ZEB1.Western Blot assay was used to detect the protien expression of ZEB1 and B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)in A549 cells.Results(1)Compared with the control group,the number of cell colonies in the sophoridine group,cisplatin group and sophoridine+cisplatin group was decreased and the apoptosis rate was increased(all P<0.05);the number of cell colonies in the sophoridine+cisplatin group was less than that in the sophoridine group and cisplatin group,and the apoptosis rate was higher than that in the sophoridine group and cisplatin group(all P<0.05).(2)Compared with the control group and the inhibitor-NC group,the levels of miR-216a-5p expression and Bax protein expression in the inhibitor group were decreased,and the mRNA and protein expression levels of ZEB1 and protein expression level of Bcl-2 were increased(all P<0.05);compared with the control grou

关 键 词：槐定碱 肺腺癌 miR-216a-5p 锌指E盒结合蛋白1 增殖 凋亡 A549细胞 

分 类 号：R285.5[医药卫生—中药学]