RESUMO
OBJECTIVE: To explore the regulatory role of miR-1908 in renal fibrosis. METHODS: The level of miR-1908 and transforming growth factor beta 1 (TGF-ß1) mRNA during renal fibrosis were detected with real-time quantitative PCR. Bioinformatics and luciferase reporter gene analyses were applied to determine the targeting relationship between miR-1908 and TGF-ß1 mRNA. After primary human renal interstitial fibroblasts were transfected with miR-1908 adenoviral expression vector in vitro, Western blotting was used to detect the protein levels of TGF-ß1, smad2/3 and matrix metalloproteinase 2 (MMP-2) in the cells. Six weeks after intraperitoneal injection of miR-1908 adenoviral vector, the renal tissue sections of the renal fibrosis mouse models were stained with Masson staining. RESULTS: Human miR-1908 showed a gradually decreasing expression during renal fibrosis process, which was completely contrary to the changes of TGF-ß1 mRNA. Overexpression of miR-1908 suppressed the expressions of TGF-ß1, smad2/3 and MMP-2 in human primary renal interstitial cells. The renal fibrosis was significantly relieved in the mice injected with miR-1908 adenovirus vector injection compared with the ones without injection. CONCLUSION: miR-1908 could inhibit renal fibrosis through targeting TGF-ß1.
Assuntos
Fibrose/metabolismo , Nefropatias/metabolismo , MicroRNAs/metabolismo , Fator de Crescimento Transformador beta1/genética , Adulto , Animais , Regulação para Baixo , Feminino , Fibrose/genética , Fibrose/patologia , Humanos , Rim/metabolismo , Rim/patologia , Nefropatias/genética , Nefropatias/patologia , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Camundongos , MicroRNAs/genética , Pessoa de Meia-Idade , Transdução de Sinais , Proteína Smad2/genética , Proteína Smad2/metabolismo , Fator de Crescimento Transformador beta1/metabolismoRESUMO
OBJECTIVE: To explore the regulatory role of CCAAT/enhancer-binding protein beta (C/EBPß) in invasion and migration of 786-O human renal cancer cells. METHODS: C/EBPß adenoviral overexpression vector (pAd-C/EBPß) and C/EBPß siRNA were constructed and respectively transfered into 786-O cells. Wound healing and Transwell(TM) invasion methods were used to detect the invasion and migration of the cells. Laser scanning confocal microscopy was applied to determine the translocation of nuclear factor κB (NF-κB), the central regulator of epithelial-mesenchymal transition (EMT); Western blotting was performed to examine the levels of EMT key markers E-cadherin, vimentin and tenascin. RESULTS: When C/EBPß was overexpressed, invasion and migration of 786-O cells were enhanced, translocation of NF-κB increased, E-cadherin level was reduced, and vimentin and tenascin levels were elevated. When C/EBPß was knocked down, the results were completely opposite to those of C/EBPß overexpression: invasion and migration of 786-O cells were inhibited, translocation of NF-κB decreased, E-cadherin level was elevated, and vimentin and tenascin expressions were suppressed. Moreover, the above changes were not found in the cells that co-treated with pAd-C/EBPß and C/EBPß siRNA. CONCLUSION: C/EBPß could promote the invasion and migration of human renal carcinoma 786-O cells, and had an inducing effect on nuclear translocation of NF-κB in 786-O cells.
