首页|LncRNA FEZF1-AS1靶向调控miR-200c-3p对人肺成纤维细胞生物学行为的影响

LncRNA FEZF1-AS1靶向调控miR-200c-3p对人肺成纤维细胞生物学行为的影响

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目的 探讨FEZ家族锌指1-反义RNA1(LncRNA FEZF1-AS1)靶向调控miR-200c-3p对人肺成纤维细胞HLF生物学行为的影响.方法 采用转化生长因子β1(TGF-β1)诱导HLF向肌成纤维细胞转化,分为空白对照组(Blank组)和造模组(HLF+TGF-β1组),另根据转染质粒不同将细胞分为Blank组、TGF-β1+Si LncRNA FEZF1-AS1 NC组和TGF-β1+Si LncRNA FEZF1-AS1组.采用Western blot法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(CollagenⅠ)和波形蛋白(Vimentin)蛋白的表达.采用实时荧光定量PCR(qRT-PCR)检测LncRNA FEZF1-AS1和miR-200c-3p的表达.采用CCK-8法检测细胞增殖,细胞划痕实验检测迁移能力,Transwell实验检测侵袭能力;采用双萤光素酶实验检测FEZF1-AS1与miR-200c-3p的靶向作用关系.结果 与Blank组比较,HLF+TGF-β1组α-SMA、CollagenⅠ、Vimentin蛋白表达及LncRNA FEZF1-AS1表达水平升高,miR-200c-3p表达水平降低(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、侵袭能力下降,LncRNA FEZF1-AS1表达及α-SMA、CollagenⅠ、Vimentin蛋白表达水平降低,miR-200c-3p表达水平升高(P<0.05);FEZF1-AS1与miR-200c-3p基因序列上存在结合位点.结论 LncRNA FEZF1-AS1通过抑制miR-200c-3p促进特发性肺间质纤维化的发生、发展.
The effect of lncRNA FEZF1-AS1 targeting regulation of miR-200c-3p on biological behaviors of human lung fibroblasts
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.

idiopathic pulmonary interstitial fibrosislung fibroblastsmyofibroblastsFEZ family zinc finger 1-antisense RNA1microRNA-200c-3p

满君、高艳艳、宋龙飞、高福生

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潍坊医学院附属医院呼吸内科(邮编 261035)

潍坊医学院附属医院康复医学科(邮编 261035)

特发性肺间质纤维化 肺成纤维细胞 肌成纤维细胞 FEZ家族锌指1-反义RNA1 微小RNA-200c-3p

国家自然科学基金资助项目

82205079

2024

天津医药
天津市医学科学技术信息研究所

天津医药

CSTPCD
影响因子:1.107
ISSN:0253-9896
年,卷(期):2024.52(3)
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