白杨素通过下调Annexin A3逆转人肺腺癌A549/DDP细胞顺铂的耐药性  

Chrysin reverses cisplatin resistance in human lung adenocarcinoma A549/DDP cells by down-regulating Annexin A3

作　　者：刘晖杰[1] 彭荣 许华[3] LIU Huijie;PENG Rong;XU Hua(Department of Oncology,Xiangyang Central Hospital Affiliated to Hubei University of Arts and Science,Xiangyang 441021,Hubei,China;Department of Oncology,Yicheng People's Hospital,Xiangyang 441021,Hubei,China;Audit Office,Xiangyang Central Hospital Affiliated to Hubei University of Arts and Science,Xiangyang 441021,Hubei,China)

机构地区：[1]湖北文理学院附属襄阳市中心医院肿瘤科,湖北襄阳441021 [2]宜城市人民医院肿瘤科,湖北襄阳441021 [3]湖北文理学院附属襄阳市中心医院审计处,湖北襄阳441021

出　　处：《贵州医科大学学报》2023年第7期800-806,共7页Journal of Guizhou Medical University

基　　金：湖北省自然科学基金(WJ2019M132)。

摘　　要：目的观察白杨素对肺腺癌顺铂(DDP)耐药性细胞A549/DDP耐药敏感性的影响。方法取A549和A549/DDP细胞,qRT-PCR及Western blot法检测膜联蛋白A3(Annexin A3)基因及蛋白表达,CCK-8法测不同浓度白杨素(0、5、25、50、100 mmoL/L)干预后细胞生存率;将A549/DDP细胞分为A549/DDP组、白杨素组(50 mol/L)、Annexin A3过表达组(Annexin A3mimic组)、Annexin A3阴性对照组(Annexin A3 NC)、白杨素+Annexin A3mimic组,另取A549细胞正常培养作为空白对照组;CCK-8法检测细胞对DDP的半数生存浓度(IC50)及耐药指数;qRT-PCR和Western blot法检测Annexin A3基因和蛋白表达水平,流式细胞义监测细胞凋亡率,小鼠荷瘤实验检测移植瘤体积。结果不同浓度的白杨素均能时效依赖性和剂量依赖性降低A549、A549/DDP细胞生存率(P<0.05),选取50 mol/L为实验浓度作用细胞48 h;与A549细胞比较,A549/DDP细胞Annexin A mRNA及蛋白表达、对DDP的IC50、耐药指数均升高,凋亡率降低(P<0.05);与A549/DDP组比较,白杨素组细胞IC50、耐药指数、Annexin A3 mRNA及蛋白表达降低,细胞凋亡率升高(P<0.05),Annexin A3 mimic组细胞IC50、耐药指数、Annexin A3 mRNA及蛋白表达升高,细胞凋亡率降低(P<0.05);Annexin A3基因高表达可减弱A549/DDP细胞对白杨素的敏感性,降低细胞凋亡率(P<0.05);小鼠荷瘤实验证实,与单独的DDP治疗组比较,白杨素与DDP联合治疗可显著降低小鼠A549/DDP移植瘤瘤体体积(P<0.05),Annexin A3基因高表达的小鼠逆转白杨素与DDP联合治疗效果(P<0.05)。结论白杨素可能通过下调Annexin A3表达,逆转A549/DDP细胞对DDP的耐药性。Objective To observe the sensitivity of lung adenocarcinoma cisplatin(DDP)resistant cell A549/DDP.Methods A549 and A549/DDP cells were collected and the gene and protein expression of Annexin A3(Annexin A3)were detected by qRT-PCR and Western blot;the cell survival rate was measured by CCK-8 method after different concentrations of chrysin(0,5,25,50,100 mmoL/L)intervention.The A549/DDP cells were divided into the A549/DDP group,chrysin group(50 mol/L),Annexin A3 overexpression group(Annexin A3mimic group),Annexin A3 negative control group(Annexin A3 NC),chrysin+Annexin A3mimic group,and normal culture of A549 cells as Blank control group.CCK-8 method measured the median survival concentration(IC50)and drug resistance index of cells in the group to DDP.qRT-PCR and Western blot methods were used to detect Annexin A3 gene and protein expression levels;flow cytometry was used to detect cell apoptosis rate;and transplanted tumor volume was detected by mouse tumor-bearing experiment.Results Different concentrations of chrysin reduced the survival rate of A549 and A549/DDP cells in a time-dependent and dose-dependent manner(P<0.05),and 50 mol/L was selected as the experimental concentration to act on the cells for 48 h.Compared with A549 cells,Annexin A mRNA and protein expression,IC50,and drug resistance index of A549/DDP cells were increased and apoptosis rate was reduced(P<0.05).Compared with the A549/DDP group,the IC50,drug resistance index,Annexin A3 gene and protein of the chrysin group were decreased,and the apoptosis rate was increased(P<0.05),and the IC50,drug resistance index,Annexin A3 gene and protein of the Annexin A3 mimic group were increased,and the apoptosis rate was reduced(P<0.05).High expression of Annexin A3 gene attenuates the sensitivity of A549/DDP cells to chrysin and reduces the rate of apoptosis.The mouse tumor-bearing experiment confirmed that compared with the DDP treatment group alone,the combined treatment of chrysin and DDP can significantly reduce the tumor volume of A549/DDP transplante

关 键 词：肺腺肿瘤 白杨素 膜联蛋白A3 顺铂耐药 A549/DDP细胞 

分 类 号：R734.2[医药卫生—肿瘤]