E2F-1靶向调控ATM激酶对鼻咽癌细胞顺铂耐药敏感性的影响  

Effect of E2F transcription factor 1 on cisplatin-resistant sensitivity of rhinitis carcinoma cells by targeting the ataxia telangiectasia mutated kinase

作　　者：贾明 边超 武凯丽 JIA Ming;BIAN Chao;WU Kaili(Department of Radiotherapy,Inner Mongolia People's Hospital,Inner Mongolia,Hohhot 010000,China)

机构地区：[1]内蒙古自治区人民医院放射治疗科,呼和浩特市010000

出　　处：《河北医药》2025年第2期188-194,共7页Hebei Medical Journal

基　　金：内蒙古自治区卫生健康委医疗卫生科技计划(编号:202102194)。

摘　　要：目的E2F转录因子1(E2F-1)靶向调控毛细血管扩张共济失调突变(ATM)激酶在顺铂(DDP)耐药鼻咽癌细胞中的作用及机制。方法选择人鼻咽癌顺铂耐药细胞系(CNE2/DDP和HNE1/DDP)及其亲代细胞(CNE2和HNE1),MTT测定细胞活力,qRT-PCR和Western blot检测E2F-1和ATM表达。将CNE2/DDP随机分为空白组(无任何处理)、对照(NC)-shRNA组(细胞用NC-shRNA质粒转染)、E2F-1 shRNA-1组(细胞用E2F-1 shRNA-1质粒转染)、E2F-1 shRNA-2组(细胞用E2F-1 shRNA-2质粒转染)、ATM组(细胞转染ATM过表达慢病毒)、E2F-1 shRNA-1+ATM组(细胞共转染E2F-1 shRNA-1质粒和ATM过表达慢病毒)、shRNA-2+ATM组(细胞共转染E2F-1 shRNA-2质粒和ATM过表达慢病毒);qRT-PCR和Western blot检测E2F-1和ATM表达,MTT检测细胞活力,qRT-PCR检测耐药和细胞周期相关基因的mRNA表达,流式细胞术检测细胞周期,EdU染色检测细胞增殖。结果与CNE2比较,CNE2/DDP细胞DDP抑制率降低,E2F-1和ATM的mRNA和蛋白表达升高(P<0.05)。与NC-shRNA组比较,E2F-1 shRNA-1组和E2F-1 shRNA-2组E2F-1和ATM的mRNA和蛋白表达降低,DDP抑制率升高,ABCA2、ABCA5、cyclin E1和CDK2表达降低,S期细胞减少,G0/G1期细胞增多,EdU阳性细胞数减少;而ATM组中ATM表达明显升高,DDP抑制率降低,ABCA2、ABCA5、cyclin E1和CDK2表达升高,S期细胞增多,G0/G1期细胞减少,EdU阳性细胞数增多(P<0.05),但2组间E2F-1的mRNA和蛋白表达差异无统计学意义(P>0.05)。与ATM组比较,E2F-1 shRNA-1+ATM组和E2F-1 shRNA-2+ATM组细胞E2F-1和ATM的mRNA和蛋白表达降低,DDP抑制率升高,ABCA2、ABCA5、cyclin E1和CDK2表达降低,S期细胞减少,G0/G1期细胞增多,EdU阳性细胞数减少(P<0.05);与E2F-1 shRNA-1组和E2F-1 shRNA-2组比较,E2F-1 shRNA-1+ATM组和E2F-1 shRNA-2+ATM组细胞ATM的mRNA和蛋白表达升高,ATM表达明显升高,DDP抑制率降低,ABCA2、ABCA5、cyclin E1和CDK2表达升高,S期细胞增多,G0/G1期细胞减少,EdU阳性细胞数增多(P<0.05),但E2F-1的mRNA和蛋白表达差�Objective To explore the role of E2F transcription factor 1(E2F-1)in cisplatin(DDP)-resistant nasopharyngeal carcinoma cells by targeting capillary dilatation ataxia telangiectasia mutated(ATM)kinase and the underlying mechanism.Methods DDP-resistant human nasopharyngeal carcinoma cell lines CNE2(CNE2/DDP)and HNE1(HNE1/DDP),and their parental cells(CNE2 and HNE1)were used to measure their cell viability by MTT assay.Expression levels of E2F1 and ATM in them were detected by the quantitative reverse transcriptase polymerase chain reaction(qRT-PCR)and Western blot.CNE2/DDP and HNE1/DDP were induced with blank control,or transfected with NC-shRNA,E2F1 shRNA-1,E2F1 shRNA-2,overexpression lentivirus of ATM,E2F1 shRNA-1+overexpression lentivirus of ATM,or E2F1 shRNA-2+overexpression lentivirus of ATM.Expression levels of E2F1 and ATM in them were detected by qRT-PCR and Western blot.The mRNA levels of genes associated with drug resistance and cell cycle progression were detected by qRT-PCR.Cell viability,cell cycle and proliferation were examined by MTT assay,flow cytometry and EdU staining,respectively.Results Compared with CNE2,CNE2/DDP had significantly lower DDP inhibition rate,but higher mRNA and protein levels of E2F1 and ATM(P<0.05).Compared with CNE2/DDP transfected with NC-shRNA,those transfected with E2F1 shRNA-1 or E2F1 shRNA-2 had significantly lower mRNA and protein levels of E2F1 and ATM,expression levels of ABCA2,ABCA5,cyclin E1 and CDK2,cells in S phase and EdU-positive cell number,but higher DDP inhibition rate and cells in G0/G1 phase(P<0.05).Compared with CNE2/DDP transfected with NC-shRNA,those transfected with overexpression lentivirus of ATM had significantly higher levels of ATM,ABCA2,ABCA5,cyclin E1 and CDK2,cells in S phase and EdU-positive cell number,but lower DDP inhibition rate and cells in G0/G1 phase(P<0.05),although mRNA and protein levels of E2F1 were comparable(P>0.05).Compared with CNE2/DDP transfected with overexpression lentivirus of ATM,those transfected with E2F1 shRNA-1+overexpre

关 键 词：鼻咽癌 E2F转录因子1 毛细血管扩张共济失调突变(ATM)激酶 顺铂耐药 

分 类 号：R739.6[医药卫生—肿瘤]