lncRNA GAS5靶向miR-21通过PI3K/Akt/mTOR信号通路抑制肺癌细胞的上皮-间质转化和自噬  

lncRNA GAS5 targets miR-21 to inhibit epithelial-mesenchymal transformation and autophagy in lung cancer cells through the PI3K/Akt/mTOR signaling pathway*

作　　者：张茜[1] 刘杰[2] 马红霞[1] ZHANG Qian;LIU Jie;MA Hongxia(Department of Respiratory and Critical Care Medicine,Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830002,China;Department of Rehabilitation,the Sixth Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830002,China)

机构地区：[1]新疆医科大学附属中医医院呼吸与危重症医学科,新疆乌鲁木齐830002 [2]新疆医科大学第六附属医院康复科,新疆乌鲁木齐830002

出　　处：《国际检验医学杂志》2025年第5期568-574,共7页International Journal of Laboratory Medicine

基　　金：“天山英才”——青年科技拔尖人才项目(2022TSYCCX0028);新疆维吾尔自治区天山创新团队计划项目(2023D14004)。

摘　　要：目的探讨长链非编码RNA(lncRNA)GAS5靶向微小RNA-21(miR-21)通过磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路抑制肺癌细胞的上皮-间质转化和自噬。方法实时定量聚合酶链反应(qPCR)检测lncRNA GAS5在5例非小细胞肺癌(NSCLC)组及癌旁组中的表达。用双荧光素酶报告基因实验检测lncRNA GAS5对miR-21的靶向作用。另将A549细胞分为5组,包括pcDNA-null组、pcDNA-GAS5组、pcDNA-GAS5+mimic-NC组、pcDNA-GAS5+mimic组、pcDNA-GAS5+mimic+BEZ235组。用qPCR检测lncRNA GAS5和miR-21表达。用蛋白质印迹法检测PI3K、磷酸化Akt(p-Akt)、磷酸化mTOR(p-mTOR)、贝克兰蛋白1(Beclin1)、微管相关蛋白1轻链3(LC3)-Ⅱ、LC3-Ⅰ、上皮钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、扭转蛋白1(Twist1)、核增殖相关抗原(Ki67)和增殖细胞核抗原(PCNA)表达。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物法检测细胞增殖活力。结果NSCLC组lncRNA GAS5表达明显高于癌旁组,差异有统计学意义(P<0.05)。双荧光素酶报告基因实验确定A549细胞中lncRNA GAS5对miR-21有直接靶向作用。与pcDNA-null组比较,pcDNA-GAS5组细胞增殖活力减少,lncRNA GAS5、PI3K、p-Akt、p-mTOR、Beclin1、LC3-Ⅱ、N-cadherin、Vimentin、Twist1、Ki67、PCNA表达下调,miR-21、LC3-Ⅰ、E-cadherin表达上调,差异有统计学意义(P<0.05)。与pcDNA-GAS5+mimic-NC组比较,pcDNA-GAS5+mimic组细胞增殖活力上调,PI3K、p-Akt、p-mTOR、Beclin1、LC3-Ⅱ、N-cadherin、Vimentin、Twist1、Ki67、PCNA表达上调,miR-21、LC3-Ⅰ、E-cadherin表达下调,差异有统计学意义(P<0.05)。与pcDNA-GAS5+mimic组比较,pcDNA-GAS5+mimic+BEZ235组细胞增殖活力下调,PI3K、p-Akt、p-mTOR、Beclin1、LC3-Ⅱ、N-cadherin、Vimentin、Twist1、Ki67、PCNA表达下调,LC3-Ⅰ和E-cadherin表达上调,差异有统计学意义(P<0.05)。结论lncRNA GAS5通过miR-21抑制PI3K/Akt/mTOR信号通路,从而Objective To investigate the inhibitory effect of long non-coding RNA(lncRNA)GAS5 on microRNA-21(miR-21)in lung cancer cells through phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway on epithalium-mesenchymal transformation and autophagy.Methods Real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of lncRNA GAS5 in 5 patients with non-small cell lung cancer(NSCLC)and adjacent groups.Dual luciferase reporter gene assay was used to detect the targeting effect of lncRNA GAS5 on miR-21.A549 cells were divided into 5 groups,including pcDNA-null group,pcDNA-GAS5 group,pcDNA-GAS5+mimic group,pcDNA-GAS5+mimic group and pcDNA-GAS5+mimic+BEZ235 group.lncRNA GAS5 and miR-21 expression were detected by qPCR.PI3K,phosphorylated Akt(p-Akt),phosphorylated mTOR(p-mTOR),Beclin1,microtubule-associated protein light chain 3(LC3)-Ⅱ,LC3-Ⅰ,E-cadherin,N-cadherin,Vimentin,Twist1,proliferation associated nuclear antigen(Ki67)and proliferating cell nuclear antigen(PCNA)expression.The cell proliferation activity was measured by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide method.Results lncRNA GAS5 expression in NSCLC group was significantly higher than that in adjacent group,with statistical significance(P<0.05).Dual luciferase reporter gene assay determined that lncRNA GAS5 in A549 cells had a direct targeting effect on miR-21.Compared with pcDNA-null group,cell proliferation activity decreased in pcDNA-GAS5 group.lncRNA GAS5,PI3K,p-Akt,p-mTOR,Beclin1,LC3-Ⅱ,N-cadherin,Vimentin,Twist1,Ki67 and PCNA were down-regulated,while miR-21,LC3-Ⅰand E-cadherin were up-regulated,the difference was statistically significant(P<0.05).Compared with pcDNA-GAS5+mimic group,the cell proliferation activity of pcDNA-GAS5+mimic group was up-regulated.The expressions of PI3K,p-Akt,p-mTOR,Beclin1,LC3-Ⅱ,N-cadherin,Vimentin,Twist1,Ki67,and PCNA were up-regulated,while the expressions of miR-21,LC3-Ⅰ,and E-cadherin were down-regulated,with s

关 键 词：长链非编码RNA GAS5 微小RNA-21 磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]