TGF-β1干预构建的人支气管上皮细胞上皮-间质转化模型中lncRNA和miRNA表达及其意义  

EXPRESSION AND SIGNIFICANCE OF LONG NON-CODING RNAS AND MICRORNAS IN EPITHELIAL-MESENCHYMAL TRANSITION MODEL OF HUMAN BRONCHIAL EPITHELIAL CELLS CONSTRUCTED BY TRANSFORMING GROWTH FACTOR-BETA 1 INTERVENTION

作　　者：陈华培 杨召川[1] 李蕾[1] 曲政海[1] CHEN Huapei;YANG Zhaochuan;LI Lei;QU Zhenghai(Pediatric Center,The Affiliated Hospital of Qingdao University,Qingdao 266555,China)

机构地区：[1]青岛大学附属医院儿科中心,山东青岛266555

出　　处：《精准医学杂志》2023年第1期32-35,共4页Journal of Precision Medicine

基　　金：国家自然科学基金青年基金项目(81700029)。

摘　　要：目的探讨人转化生长因子β1(TGF-β1)干预构建的人支气管上皮细胞16HBE上皮-间质转化(EMT)模型中lncRNA和miRNA的表达情况。方法体外培养16HBE细胞,设立对照组和处理组。处理组以TGF-β1诱导细胞构建EMT细胞模型,对照组加入等量RPMI基础培养液。分别于TGF-β1处理细胞后24、48、72 h使用倒置显微镜观察两组细胞形态及分布。当处理组细胞开始出现明显梭形改变时,采用RNA分离、逆转录以及实时荧光定量PCR(RT-qPCR)方法检测两组细胞中lncRNA和miRNA相对表达量。结果经TGF-β1处理细胞48 h后,处理组细胞梭形改变明显,细胞间隙增大,显示EMT模型构建成功。同时在该时间点,两组细胞中TBILA、NKILA、LNCRNA-ATB、HOTAIR、NEAT 1、miR-125b-5p、miR-21-3p、miR-21-5p、miR-27a-3p、miR-27a-5p、miR-141-3p、miR-200c-3p、miR-17-5p、miR-34a-5p的相对表达量比较差异均具有显著意义(t=-16.353~6.460,P<0.05)。结论TGF-β1干预能够成功构建EMT细胞模型,该细胞模型中的细胞与正常细胞相比,多个lncRNA和miRNA表达具有显著差异。上述基因可能参与了EMT或气道重塑的形成和发展,或许为这些疾病的防治提供分子理论支持。Objective To investigate the expression of long non-coding RNAs(lncRNAs)and microRNAs(miRNAs)in the epithelial-mesenchymal transition(EMT)model of human bronchial epithelial cell line 16HBE induced by human transforming growth factor-β1(TGF-β1).Methods 16HBE cells were cultured in vitro,and the experiment established control group and treatment group.The cells in the treatment group were induced by TGF-β1 to establish an EMT cell model,and those in the control group were added with an equal volume of RPMI basic culture medium.At 24,48,and 72 h after TGF-β1 treatment,an inverted microscope was used to observe cell morphology and distribution.At the time point when the cells in the treatment group began to show obvious spindle-shaped changes,the methods of RNA separation,reverse transcription,and quantitative real-time PCR were used to measure the relative expression levels of lncRNAs and miRNAs in the two groups.Results After 48 h of TGF-β1 treatment,the cells in the treatment group showed obvious spindle-shaped changes and an increase in intercellular space,suggesting that the EMT model was successfully established.At this time point,there were significant differences between the two groups in the relative expression levels of TBILA,NKILA,LNCRNA-ATB,HOTAIR,NEAT 1,miR-125b-5p,miR-21-3p,miR-21-5p,miR-27a-3p,miR-27a-5p,miR-141-3p,miR-200c-3p,miR-17-5p,miR-34a-5p(t=-16.353-6.460,P<0.05).Conclusion TGF-β1 intervention can successfully establish the EMT cell model,and the differentially expressed lncRNAs and miRNAs between this cell model and normal cells may be involved in the formation and development of EMT or airway remodeling,which might provide a theoretical basis for the prevention and treatment of such diseases.

关 键 词：RNA 长链非编码 微RNAS 转化生长因子β1 上皮-间质转化 上皮细胞 气道重塑 纤维化 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R394[医药卫生—基础医学]