miR-138/sirt-l调控非小细胞肺癌增殖和转移的机制研究  

Mechanism of miR-138/sirt-l axis on proliferation and metastasis of non-small cell lung cancer

作　　者：朱冬伊 卢青纯 李占霞[1] 顾莉 任涛[1] Zhu Dongyi;Lu Qingchun;Li Zharucia;Gu Li;Ren Tao(Department of Respiratory Medicitie,Shanghai East Hospital,Tongji Vniuersity School ofMedicine,Shanghai 200120,China)

机构地区：[1]同济大学附属东方医院呼吸内科,上海200120

出　　处：《国际呼吸杂志》2019年第4期249-256,共8页International Journal of Respiration

摘　　要：目的探讨微小RNA138(miR-138)/沉默信息调节因子1(sirt-1)调控非小细胞肺癌(NSCLC)增殖和转移的机制研究。方法反转录实时定量聚合酶链式反应(RT-qPCR)检测sirt-1在临床样本NSCLC组织和癌旁组织的表达。选取NSCLC细胞株A549和H1299,通lipo3000分别转染空质粒、miR-138成熟体和miR-138抑制剂.将实验分组为对照组、miR-138组、miR-138抑制组。双荧光素酶报告、RT-qPCR和蛋白质印迹法(Western blot)检测miR-138与sirt-1的靶向关系;Cell Counting Kit-8,EdU检测miR-138对A549和H1299增殖的影响;划痕实验、Transwell检测miR-138对A549和H1299迁移和侵袭的影响;Western blot检测上皮间叶转化标记物[上皮-钙黏素(E-cadherin)、波形蛋白(Vimentin)]的表达。结果sirt-1 mRNA水平在NSCLC组织较癌旁组织表达明显升高;双荧光素酶报告提示共转miR-138成熟体+野生型sirt-1质粒较其他质粒活性减低;与对照组比较,miR-138组中sirt-1表达下降,miR-138抑制组中sirt-1表达升高;与对照组比较.miR-138组中细胞增殖和转移降低,miR-138抑制组的细胞增殖和转移升高;与对照组比较,miR-138组A549中E-cadherin表达升高,两细胞株vimentin表达下降,而miR-138抑制后,结果完全相反’结论miR-138通过调控sirt-1抑制NSCLC的增殖和转移,提示miR-138/sirt-l可能成为临床治疗NSCLC潜在位点。Objective To investigate the mechanism of microRNA138 ( miR-138)/silence information regulatorl ( sirt-1 ) on proliferation and metastasis of non-small cell lung cancer (NSCLC). Methods Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expression of sirt-1 on NSCLC tissues and adjacent tissues. After NSCLC cell lines A549 and H1299 were selected and transfected with empty plasmid, miR-138 mimics and miR- 138 inhibitor by lipo3000, the experiment was divided into negative control group, miR-138 group and miR-138 inhibition group. The relation between miR-138 and sirt-1 was evaluated by dual luciferase reporter assay, RT-qPCR and Western blot;the effect of miR-138 on the proliferation of A549 and H1299 were detected by Cell Counting Kit-8 and EdU;the effect of miR-138 on the migration and invasion was detected by Wound-healing and Transwell assays;the expression of epithelial-mesenchymal transition markers including E-cadherin and Vimentin were detected by Western blot. Results The expression of sirt-1 on mRNA level was obviously increased in NSCLC tissues, compared to adjacent tissues. Dual luciferase reporter assay showed the luciferase activity on cotransfection of miR-138 mimics+ sirt-1 wildtype plasmid was lower than others. The expression of sirt-1 in miR-138 group was decreased and that in miR-138 inhibition group was increased, compared to negative control group. The proliferation and metastasis of NSCLC cells in miR-138 group were decreased and those in miR-138 inhibition group were increased, compared to negative control group. The expression of E-cadherin in A549 was increased and the expression of Vimentin in both cell lines were decreased in miR-138 group, compared to negative control group. When miR 138 was inhibited, the result was completely opposite. Conclusions miR-138 suppresses the proliferation and metastasis of NSCLC by regulating sirt-1, which indicates that miR-138 sirt-1 might be a potential therapeutic target for NSCLC.

关 键 词：沉默信息调节因子1 上皮间质转化 微小RNA138 癌 非小细胞肺 

分 类 号：R734.2[医药卫生—肿瘤]