摘要
目的 探讨FEZ家族锌指1-反义RNA 1(LncRNA FEZF1-AS1)对肺间质纤维化细胞增殖、迁移、侵袭能力及上皮-间充质相互作用(EMT)的影响及其作用机制.方法 将转化生长因子-β1(TGF-β1)作用于A549细胞,诱导肺间质纤维化细胞模型,将其分为空白对照组(A549细胞组)和模型组.Western blotting检测细胞E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)及波形蛋白(Vimentin)表达,观察模型复制是否成功;实时荧光定量聚合酶链反应(qRT-PCR)检测细胞LncRNA FEZF1-AS1和miR-200c-3p基因表达.根据实验目的和转染质粒不同,将A549细胞分为空白对照组(Blank组)、TGF-β1+Si LncRNA FEZF1-AS1 NC组、TGF-β1+Si LncRNA FEZF1-AS1组.CCK-8法检测细胞增殖能力;细胞划痕实验检测细胞迁移能力;Transwell检测细胞侵袭能力.Western blotting检测细胞E-cadherin、N-cadherin及Vimentin蛋白表达;qRT-PCR检测细胞LncRNA FEZF1-AS1和miR-200c-3p基因表达.结果 模型组E-cadherin蛋白相对表达量较空白对照组降低(P＜0.05),N-cadherin及Vimentin蛋白相对表达量较空白对照组升高(P＜0.05);模型组LncRNA FEZF1-AS1 基因相对表达量较空白对照组升高(P＜0.05),miR-200c-3p 基因相对表达量较空白对照组降低(P＜0.05).与Blank组比较,TGF-β1+Si LncRNA FEZF1-AS1 NC组细胞增殖、迁移、侵袭能力升高(P＜0.05);与TGF-β1+Si LncRNA FEZF1-AS1 NC组比较,TGF-β1+Si LncRNA FEZF1-AS1组细胞增殖、迁移、侵袭能力降低(P＜0.05).与TGF-β1+Si LncRNA FEZF1-AS1 NC组比较,TGF-β1+Si LncRNA FEZF1-AS1组LncRNA FEZF1-AS1基因相对表达量及N-cadherin、Vimentin蛋白相对表达量降低(P＜0.05),E-cadherin蛋白相对表达量升高(P＜0.05);与TGF-β1+Si LncRNA FEZF1-AS1 NC组比较,TGF-β1+Si LncRNA FEZF1-AS1 组和Blank组miR-200c-3p 基因相对表达量升高(P＜0.05).结论 LncRNA FEZF1-AS1通过抑制miR-200c-3p促进肺间质纤维化细胞增殖、迁移、侵袭及EMT过程.
Abstract
Objective To investigate the effects of the long non-coding RNA FEZF1-AS1(LncRNA FEZF1-AS1)on the proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)of cells in idiopathic pulmonary fibrosis(IPF)and its underlying mechanisms.Methods A549 cells were treated with Transforming growth factor-β1(TGF-β1)to induce a pulmonary fibrosis cell model,dividing them into a blank control group(A549 cell group)and a model group.Western blotting was used to measure the expression of E-cadherin,N-cadherin,and Vimentin to verify the model's success.Real-time quantitative polymerase chain reaction(qRT-PCR)was employed to measure the expression of LncRNA FEZF1-AS1 and microRNA-200c-3p(miR-200c-3p).Based on experimental goals and different transfection plasmids,A549 cells were divided into three groups:Blank,TGF-β1+Si LncRNA FEZF1-AS1 NC,and TGF-β1+Si LncRNA FEZF1-AS1.Cell proliferation was evaluated using the CCK-8 assay,migration was assessed with wound healing assays,and invasion capabilities were measured using Transwell assays.Western blotting and qRT-PCR were repeated to assess protein and gene expression changes.Results In the model group,the relative expression of E-cadherin decreased,while N-cadherin and Vimentin increased compared to the blank control(P＜0.05).LncRNA FEZF1-AS1 was upregulated,and miR-200c-3p was downregulated in the model group compared to the control(P＜0.05).Compared to the Blank group,cell proliferation,migration,and invasion were increased in the TGF-β1+Si LncRNA FEZF1-AS1 NC group(P＜0.05).In contrast,these parameters were reduced in the TGF-β1+Si LncRNA FEZF1-AS1 group compared to the NC group(P＜0.05).Also,this group showed decreased expression of LncRNA FEZF1-AS1,N-cadherin,Vimentin,and increased E-cadherin compared to the NC group(P＜0.05).The expression of miR-200c-3p was higher in the TGF-β1+Si LncRNA FEZF1-AS1 and Blank groups compared to the NC group(P＜0.05).Conclusions LncRNA FEZF1-AS1 promotes the proliferation,migration,invasion,and EMT processes in pulmonary fibrosis cells by inhibiting miR-200c-3p.
维普
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