Epithelial-mesenchymal transition(EMT)is a vital pathological feature of silica-induced pulmonary fibrosis.However,whether circRNA is involved in the process remains unclear.The present study aimed to investigate the ...Epithelial-mesenchymal transition(EMT)is a vital pathological feature of silica-induced pulmonary fibrosis.However,whether circRNA is involved in the process remains unclear.The present study aimed to investigate the role of circPVT1 in the silica-induced EMT and the underlying mechanisms.We found that an elevated expression of circPVT1 promoted EMT and enhanced the migratory capacity of silica-treated epithelial cells.The isolation of cytoplasmic and nuclear separation assay showed that circPVT1 was predominantly expressed in the cytoplasm.RNA immunoprecipitation assay and RNA pull-down experiment indicated that cytoplasmic-localized circPVT1 was capable of binding to miR-497-5p.Furthermore,we found that miR-497-5p attenuated the silica-induced EMT process by targeting transcription factor 3(TCF3),an E-cadherin transcriptional repressor,in the silica-treated epithelial cells.Collectively,these results reveal a novel role of the circPVT1/miR-497-5p/TCF3 axis in the silica-induced EMT process in lung epithelial cells.Once validated,this finding may provide a potential theoretical basis for the development of interventions and treatments for pulmonary fibrosis.展开更多
目的探讨miR-497-5p对子宫内膜癌Ishikawa细胞侵袭、迁移和上皮间质转化(EMT)的影响及其机制。方法将体外培养的Ishikawa细胞分为miR-NC组、miR-497-5p组、pLV-VEGFA组和miR-497-5p+VEGFA组,采用Real time PCR检测miR-497-5p和血管内皮...目的探讨miR-497-5p对子宫内膜癌Ishikawa细胞侵袭、迁移和上皮间质转化(EMT)的影响及其机制。方法将体外培养的Ishikawa细胞分为miR-NC组、miR-497-5p组、pLV-VEGFA组和miR-497-5p+VEGFA组,采用Real time PCR检测miR-497-5p和血管内皮生长因子A(VEGFA)mRNA的表达,双荧光素酶报告基因实验检测miR-497-5p和VEGFA的靶向关系,Western blot检测VEGFA蛋白和EMT相关蛋白神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和E-钙黏蛋白(E-cadherin)的表达,Transwell小室实验检测Ishikawa细胞的侵袭和迁移。结果转染miR-497-5p mimics可升高Ishikawa细胞中miR-497-5p表达,降低VEGFA mRNA表达(P<0.05),VEGFA是miR-497-5p的靶基因。与miR-NC组比较,miR-497-5p组细胞中VEGFA、Vimentin和E-cadherin蛋白的表达水平、侵袭细胞数和迁移细胞数均明显降低,而N-cadherin蛋白表达升高(P<0.05);与pLV-VEGFA组比较,miR-497-5p+VEGFA组细胞中VEGFA、Vimentin和E-cadherin蛋白的表达水平以及侵袭、迁移细胞数量均明显降低,而N-cadherin蛋白表达明显升高(P<0.05)。结论miR-497-5p可靶向VEGFA抑制子宫内膜癌Ishikawa细胞的侵袭、迁移和上皮间质转化。展开更多
基金funded by the National Natural Science Foundation of China(Grant No.82073518).
文摘Epithelial-mesenchymal transition(EMT)is a vital pathological feature of silica-induced pulmonary fibrosis.However,whether circRNA is involved in the process remains unclear.The present study aimed to investigate the role of circPVT1 in the silica-induced EMT and the underlying mechanisms.We found that an elevated expression of circPVT1 promoted EMT and enhanced the migratory capacity of silica-treated epithelial cells.The isolation of cytoplasmic and nuclear separation assay showed that circPVT1 was predominantly expressed in the cytoplasm.RNA immunoprecipitation assay and RNA pull-down experiment indicated that cytoplasmic-localized circPVT1 was capable of binding to miR-497-5p.Furthermore,we found that miR-497-5p attenuated the silica-induced EMT process by targeting transcription factor 3(TCF3),an E-cadherin transcriptional repressor,in the silica-treated epithelial cells.Collectively,these results reveal a novel role of the circPVT1/miR-497-5p/TCF3 axis in the silica-induced EMT process in lung epithelial cells.Once validated,this finding may provide a potential theoretical basis for the development of interventions and treatments for pulmonary fibrosis.