Résumé
Objective To investigate the effects of long non-coding RNA FEZ family zinc finger 1 antisense RNA 1(lncRNA FEZF1-AS1)on enhancer of zeste homolog 2(EZH2)in regulation of proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of pulmonary interstitial cells and its mechanism.Methods The A549 cells human lung adenocarcinoma cell line were divided into control group and model group[model cells were induced into lung interstitial cells after being treated with transforming growth factor β1(TGF-β1)20 ng/mL for 48 h].The protein expression of E-cadherin,N-cadherin and vimentin in each group was detected by Western blot.The expression of lncRNA FEZF1-AS1 and EZH2 in the two groups was detected by RT-qPCR.Cells in the trans-fection group were divided into si NC group,lncRNA FEZF1-AS1+OE vector group and si lncRNA FEZF1-AS1+OE EZH2 group.Cell proliferation was examined by CCK-8 method,cell migration was detected by cell scratch,and cell invasion was detected by Transwell assays.The protein expression of E-cadherin,N-cadherin,vimentin and EZH2 in each group was detected by Western blot.The direct binding effect of FEZF1-AS1 and EZH2 was deter-mined by RNA immuno-precipitation(RIP).Results Compared with the control group,the protein expression level of E-cadherin in the model group was significantly decreased(P＜0.05),and the protein expression of N-cadherin and vimentin was significantly increased(P＜0.05).Compared with the control group,the expression level of lncRNA FEZF1-AS1 and EZH2 genes was significantly increased in the model group(P＜0.05).Compared with si NC group,the proliferation,migration and invasion ability of si lncRNA FEZF1-AS1+OE vector group were decreased,the ex-pression of E-cadherin protein was increased while the expression of N-cadherin,vimentin and EZH2 was decreased(P＜0.05).Compared with si lncRNA FEZF1-AS1+OE vector group,the proliferation,invasion and migration of si lncRNA FEZF1-AS1+OE EZH2 group were increased(P＜0.05).E-cadherin expression was decreased,while N-cad-herin,vimentin and EZH2 expressions were increased(P＜0.05).RIP experiment further confirmed that lncRNA FEZF1-AS1 had direct binding effect with EZH2.Conclusions LncRNA FEZF1-AS1 can promote the proliferation,invasion,metastasis and EMT process of pulmonary fibrosis cells by regulating EZH2.
Résumé
Objective To investigate the effect of FEZ family zinc finger 1-antisense RNA 1(LncRNA FEZF1-AS1)targeting regulation of miR-200c-3p expression on biological behaviors of human lung fibroblasts(HLF).Methods Transforming growth factor β1(TGF-β1)was used to induce the transformation of HLF into myofibroblasts,which were divided into the Blank group and the model group(HLF+TGF-β1 group).According to different transfection plasmid,cells were divided into the Blank group,the TGF-β1+Si LncRNA FEZF1-AS1 NC group and the TGF-β1+Si LncRNA FEZF1-AS1 group.The protein expressions of α-SMA,Collagen Ⅰ and Vimentin were detected by Western blot assay.The expressions of LncRNA FEZF1-AS1 and miR-200c-3p were detected by quantitative real-time PCR(qRT-PCR).Cell proliferation ability was detected by CCK-8 method,migration ability was detected by cell scratch experiment and invasion ability was detected by Transwell assay.The targeting relationship between FEZF1-AS1 and miR-200c-3p was detected by dual-luciferase reporter assay.Results Compared with the Blank group,protein expressions of α-SMA,Collagen Ⅰ,Vimentin and the expression of LncRNA FEZF1-AS1 were increased in the HLF+TGF-β1 group(P＜0.05),and the expression of miR-200c-3p was decreased(P＜0.05).Compared with the TGF-β1+Si LncRNA FEZF1-AS1 NC group,cell proliferation,migration,invasion ability,LncRNA FEZF1-AS1 expression,protein expressions of α-SMA,Collagen Ⅰ and Vimentin were decreased in the TGF-β1+Si LncRNA FEZF1-AS1 group(P＜0.05),and the expression of miR-200c-3p was increased(P＜0.05).There were binding sites between miR-200c-3p and FEZF1-AS1 gene sequence.Conclusion LncRNA FEZF1-AS1 promotes the formation and progression of idiopathic pulmonary interstitial fibrosis by inhibiting miR-200c-3p.