miR-34a通过激活AMPK/mTOR通路增强鼻咽癌细胞顺铂敏感性  

miR-34a enhance the cisplatin sensitivity of nasopharyngeal carcinoma cells by activating AMPK/mTOR pathway

作　　者：李雪娟[1] 马菲[1] 马庆华[1] LI Xuejuan;MA Fei;MA Qinghua(Department of Otorhinolaryngologic,Zhengzhou Seventh People's Hospital,Henan Zhengzhou 450000,China)

机构地区：[1]郑州市第七人民医院耳鼻喉科,河南郑州450000

出　　处：《现代肿瘤医学》2022年第3期402-406,共5页Journal of Modern Oncology

基　　金：河南省医学科技攻关计划联合共建项目(编号:LHGJ20190046)。

摘　　要：目的:探讨微小RNA(microRNA,miR)-34a对鼻咽癌细胞顺铂敏感性的影响及其机制。方法:采用实时荧光定量PCR法检测鼻咽癌顺铂耐药HNE1/DDP细胞和亲本HNE1细胞中miR-34a的表达水平。将HNE1/DDP细胞分为未转染组(正常培养)、miR-NC组(转染阴性对照)和miR-34a组(转染miR-34a模拟物),实时荧光定量PCR法检测细胞中miR-34a表达,噻唑蓝[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]法检测细胞对顺铂的敏感性,流式细胞仪检测细胞凋亡率,免疫印迹法检测凋亡相关蛋白B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)及腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路相关蛋白AMPK、磷酸化(p)-AMPK、mTOR、p-mTOR的表达水平。结果:与HNE1细胞比较,HNE1/DDP细胞中miR-34a表达水平明显降低(P<0.05);与miR-NC组比较,miR-34a组细胞中miR-34a表达水平、细胞凋亡率、Bax蛋白表达水平及p-AMPK/AMPK水平明显升高,顺铂对细胞的半数抑制浓度(the half inhibitory concentration,IC_(50))、Bcl-2蛋白表达水平和p-mTOR/mTOR水平均明显降低(P<0.05)。结论:miR-34a过表达可通过诱导细胞凋亡增强鼻咽癌HNE1/DDP细胞顺铂敏感性,其作用机制可能与激活AMPK/mTOR通路有关。Objective:To investigate the effect of microRNA(miR)-34a on cisplatin sensitivity of nasopharyngeal carcinoma cells and its mechanism.Methods:The expression level of miR-34a in cisplatin resistant HNE1/DDP cells and parent HNE1 cells was detected by real-time fluorescent quantitative PCR.HNE1/DDP cells were divided into non-transfection group(normal culture),miR-NC group(transfection negative control)and miR-34a group(transfection of miR-34a mimic),the expression of miR-34a was detected by real-time fluorescent quantitative PCR,the sensitivity of cells to cisplatin was detected by methyl thiazolyl tetrazolium(MTT)assay,the apoptosis rate was detected by flow cytometry,Western blotting was used to detect the expression levels of apoptosis associated proteins B-lymphoma-2 gene(Bcl-2),Bcl-2-related X protein(Bax),and AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)pathway proteins AMPK,phosphorylation(p)-AMPK,mTOR and p-mTOR.Results:Compared with that of HNE1 cells,the expression level of miR-34a in HNE1/DDP cells was significantly lower(P<0.05).Compared with that in miR-NC group,miR-34a expression level,apoptosis rate,Bax protein expression level and p-AMPK/AMPK level in miR-34a group were significantly higher,the half inhibitory concentration of cisplatin on cells(IC_(50)),Bcl-2 protein expression level and p-mTOR/mTOR level were significantly lower(P<0.05).Conclusion:Overexpression of miR-34a can enhance the sensitivity of HNE1/DDP cells to cisplatin by inducing apoptosis,which may be related to the activation of AMPK/mTOR pathway.

关 键 词：鼻咽癌 微小RNA-34a 顺铂 腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白通路 

分 类 号：R739.6[医药卫生—肿瘤]