西安交通大学学报（医学版）2017，Vol.38Issue(2)：176-181,6.DOI:10.7652/jdyxb201702005
TTF-1启动子调控miR-7表达对人肺癌95D细胞体外凋亡的影响
Effect of TTF-1 promoter regulated miR-7 expression on the apoptosis of human lung cancer 95D cells in vitro
摘要
Abstract
Objective To investigate the effect of thyroid transcription factor-1 (TTF-1)promoter regulated miR-7 expression on the apoptosis of human lung cancer 95D cells invitro and to explore its significance.Methods A eukaryotic expression vector of TTF-1 was detected by FCM,and the apoptosis of cells was analyzed by TUNEL assay.Furthermore,the expression levels of phos-Casp-3 and phos-Casp-9 protein in 95D cells were detected by Western blot.Meanwhile,the expression of CGGBP1 and EGFR,twotarget molecules of miR-7,were examined by immunofluorescence assay confocal microscopy. Finally, the expression of phosphorylated Akt and promoter regulation miR-7 (p-TmiR-7 ) and control vector p-Cont which had been previous constructed were transiently transfected into the human lung cancer 95D cells invitro,respectively.TTF-1 promoter regulated miR-7 eukaryotic expression vector (named p-TmiR-7)and control vector p-Cont were transiently transfected into human lung cancer 95D cells in vitro,respectively.The expressions of phosphorylated Akt and Erk protein in each group were assayed by Western blot.Results At 48 h after transfection invitro,the apoptosis ratio of cells in p-T-miR-7 transfection group significantly increased compared with that in p-Cont transfection group (P <0.05 ).Meanwhile,the expression levels of phos-Casp-3 and phos Casp-9 protein significantly increased (P<0.05 ).Furthermore,the expressions of EGFR and CGGBP1 protein decreased obviously,accompanied by decreased expressions of phos-Akt and phos-Erk in p-T-miR-7 transfection group (P<0.05 ).Conclusion TTF-1 promoter regulated miR-7 expression can induce the apoptosis of human lung cancer cells in vitro,which provides some experimental basis for the development of human lung cancer gene targeting therapeutic strategy.关键词
miR-7/甲状腺转录因子-1(TTF-1)/启动子/基因治疗/肺癌Key words
miR-7/thyroid transcription factor-1 (TTF-1 )/promoter/gene therapy/lung cancer分类
医药卫生引用本文复制引用
卢佳,陈超,廖珍媛,崔盼盼,雷良玉,徐华林,田丹,郑静,徐林..TTF-1启动子调控miR-7表达对人肺癌95D细胞体外凋亡的影响[J].西安交通大学学报（医学版）,2017,38(2):176-181,6.基金项目
国家自然科学基金资助项目(No.31370918),贵州省高层次人才项目(黔科合人才(2016)4031 号),贵州省教育厅特色重点实验室建设项目(黔教合KY字[2014]212) Supported by the National Natural Science Foundation of China (No.31370918),Guizhou Provincial High-level Talents Program [No.(2016) 4031],and Characteristic Key Laboratory Construction Program of Guizhou Provincial Department of Education [No.(2014)212] （No.31370918）
