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PI3K/AKT途径在Gli诱导的肺鳞癌SK-MES-1细胞上皮-间质转化中的作用

Role of PI3K/AKT pathway in Gli promoting epithelial-mesenchymal transition in lung squamous carcinoma SK-MES-1 cell
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摘要 目的 研究Gli与磷酯酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)诱导的肺鳞癌SK-MES-1细胞上皮-间质转化之间的关系.方法 实时荧光定量聚合酶链反应(PCR)法观察GANT61处理SK-MES-1细胞24h后对Gli1、Gli2、波形蛋白(Vimentin)和钙黏附蛋白E(E-cadherin)mRNA表达的影响;Western blot试验观察GANT61和N-Shh分别处理SK-MES-1细胞24h后对Gli1、Gli2、p-AKT、AKT、Vimentin和E-cadherin蛋白表达的影响;侵袭实验观察对肺鳞癌细胞迁移侵袭能力的影响.结果 GANT61与二甲基亚砜(DMSO)相比可以显著下调肺鳞癌SK-MES-1细胞的Gli1、Gli2、Vimentin mRNA表达,上调E-cadherin mRNA的表达(P均〈0.01);Western blot显示GANT61可以下调SK-MES-1的Gli1、Gli2、p-AKT、Vimentin蛋白表达,上调E-cadherin蛋白表达(P值均〈0.01).侵袭实验显示GANT61可以显著抑制SK-MES-1细胞的侵袭能力(P〈0.01).应用N-Shh可以显著上调Gli1、Gli2、Vimentin mRNA及蛋白表达以及p-AKT蛋白表达,下调E-cadherin mRNA和蛋白的表达,同时促进肿瘤细胞的侵袭能力.结论 Gli1和Gli2表达下调可以抑制肺鳞癌细胞的侵袭转移,可能与Gli通过PI3K/AKT途径促进上皮-间质转化有关. Objective To explore the molecular mechanisms that Gli and PI3K/AKT regulated epithelial-mesenchymal transition in lung squamous carcinoma . Methods Real time fluorescence quantitative polymerase chain reaction ( PCR) method explained the mRNA expression of Gli1、Gli2、Vimentin and E-cadherin after cells were treated by GANT 61 in lung squamous carcinoma .Western blot assay observed the protein expression of Gli1 , Gli2 , p-AKT , AKT , Vimentin and E-cadherin with treatment of GANT61 .Cell invasion ability was detected by transwell assay .Meanwhile ,N-Shh was applied to activate the Hedgehog pathway in respective .Results Compared with DMSO ,GANT61 could significantly down-regulate the mRNA expression of Gli1 ,Gli2 ,Vimentin and up-regulate E-cadherin mRNA(all P 〈 0 .01) ,Western blot assay displayed the fact that GANT61 could down-regulate Gli1 , Gli2 ,p-AKT ,Vimentin protein expression and up-regulate E-cadherin protein expression ( all P 〈0 .01) , GANT61 could obviously inhibit the ability of invasion in SK-MES-1 by transwell assay ( P 〈0 .01) .N-Shh proteins which activated Hedgehog pathway could up-regulate Gli1 ,Gli2 ,Vimentin mRNA and Gli1 , Gli2 ,Vimentin and p-AKT protein expression ,meanwhile down-regulate E-cadherin mRNA and protein expression ,and increase cell invasion ability . Conclusions Down-regulate Gli1 and Gli2 can inhibit metastasis of lung squamous carcinoma ,and it may be related to promoting epithelial-mesenchymal transition through PI3K/AKT pathway .
作者 杜媛鲲 米源 陈阁 廖海江 王雷 Du Yuankun , Mi Yuan, Chen Ge, Liao Haijiang, Wang Lei. ( Department of Periodical Magazine, Hebei Medical University, Shijiazhuang 050017, China)
出处 《国际呼吸杂志》 2017年第24期1901-1906,共6页 International Journal of Respiration
基金 河北省引进留学人员资助项目(CY201613)
关键词 肺鳞癌 Hedgehog/Gli P13K/AKT GANT61 上皮-间质转化 Lung squamous carcinoma Hedgehog/Gli PI3K/AKT GANT61 Epithelial-Mesenchymal Transition
作者简介 通信作者:王雷,Email:492850006@qq.com
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