The Effect And Mechanism Of MicroRNA-127 Onthe Progression Of Lung Cancers

Objective:To investigate the role of mi R-127 in NSCLC and its molecular mechanism.Methods:1.Determination mi R-127 in lung cancer tissue and cell lines by Real time PCR and in situ hybridization; Real time PCR to determinate the efficiency of PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line.2.Determination EMT relative molecular in PC9 overexpression miR-127 cell line and A549 konckdown mi R-127 cell line by Real time PCR and Western blot.3.Determination migration and invasion in PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line by scratch test and Transwell.4.Determination p-ERK、p-AKT and apoptosis in PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line treated Gefitinib by MTS and Western blot. To establish an subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice. Determination Vimentin 、 K i67 、 MMP9 and TNFAIP3 by immunohistochemistry.5.Determination TNFAIP3 in PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line by Real time PCR and Western blot. Luciferase activity of TNFAIP3 3’UTR was analyzed in PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line.6.Determination p-IKKα/β, p-IκBα, p-p65 and RIP1 K63 ubiquitin in PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line treated TNF-α by Western blot.7.Determination TNFAIP3 resume the regulation of mi R-127 by scratch test,Transwell, MTS and Western blot.Results:1.mi R-127 is overexpression in lung cancer tissues and cell lines.2.PC9 overexpression of mi R-127 caused significant increase of N-Cadherin、ZEB1 and Vimentin, down-regulation of E-Cadherin. A549 konckdown mi R-127 down-regulation of N-Cadherin, ZEB1 and Vimentin, increase of E-Cadherin.3.mi R-127 increased migration and invasion in PC9 overexpression mi R-127 cell line; Anti- mi R-127 down-regulation migration and invasion in A549 konckdown mi R-127 cell line.4.mi R-127 promoted the lung cancer cell growth in vitro and vivo.Gefitinib increased AKT and ERK activation. PC9 overexpression of mi R-127 treated Gefitinib caused significant increase of p-ERK、AKT and the tumorogenesis. A549 konckdown mi R-127 down-regulation of p-ERK, AKT. mi R-127 increased MMP9、K i67 and Vimentin; mi R-127 down-regulation of TNFAIP3.5.mi R-127 repressed TNFAIP3 in PC9 overexpression mi R-127 cell line; Anti- mi R-127 increased TNFAIP3 in A549 konckdown mi R-127 cell line. Luciferase activity of TNFAIP3 3’UTR was analyzed in PC9 overexpression mi R-127 cell line and A549 konckdown mi R-127 cell line.6.PC9 overexpression of mi R-127 caused significant increase of p-IKKα/β, p-IκBα, p-p65 and RIP1 K63 ubiquitin. A549 konckdown mi R-127 down-regulation of p-IKKα/β, p-IκBα, p-p65 and RIP1 K63 ubiquitin. TNFAIP3 could resume the process.7. TNFAIP3 resume the regulation of mi R-127.Conclusions:1.mi R-127 was overexpression in non-small-cell lung cancer(NSCLC).2.mi R-127 caused induction of epithelial- to-mesenchymal transition(EMT), growth, cell migration and invasion in NSCLC.3. miR-127 promoted the resistance of Gefitinib.4.TNFAIP3 was a molecular target of miR-127 and actived NF-κB by inhibiting TNFAIP3 in NSCLC.5. TNFAIP3 resume the regulation of mi R-127 in NSCLC.