Karetnikova Ekaterina Sergeevna, Jarzebska Natalia, Rodionov Roman Nikolaevich, Rubets Elena, Markov Alexander Georgievich, Spieth Peter Markus
Department of Anesthesiology and Critical Care Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
Department of General Physiology, Saint-Petersburg State University, Saint-Petersburg, Russia.
Rep Biochem Mol Biol. 2023 Jul;12(2):211-219. doi: 10.61186/rbmb.12.2.211.
Epithelial-mesenchymal transition (EMT) is an important physiologic process that determines the outcome of lung tissue healing after injury. Stimuli and molecular cascades inducing EMT lead to up-regulation of the mesenchymal-specific genes in the alveolar epithelial cells and to down-regulation of the genes coding for epithelial markers. Alveolar epithelial cell lines are commonly used as in vitro models to study processes occurring in the lung tissue. The aim of this study is to quantify and compare mRNA expression levels of epithelial and mesenchymal markers in a number of lung epithelial cell lines.
Lung epithelial cell lines L2, R3/1 and RLE-6TN were cultured. Repeated mRNA isolation, reverse transcription, and quantitative PCR with primers to epithelial (E-cadherin, occludin, and ZO-2) and mesenchymal (α-SMA, collagen III, and vimentin) markers were performed.
First, our study revealed a higher level of epithelial transcripts in the RLE-6TN cell line compared to L2 and R3/1 cells. Secondly, we have found simultaneous mRNA expression of both epithelial (E-cadherin, occludin and ZO-2) and mesenchymal (α-SMA, collagen III and vimentin) markers in all cell lines studied.
Our data indicate that at the transcriptional level the L2, R3/1, and RLE-6TN cell lines are at one of the intermediate stages of EMT, which opens new possibilities for the study of EMT on cell lines. Determination of the direction of changes in epithelial and mesenchymal markers will make it possible to establish the factors responsible for both EMT and reverse mesenchymal-epithelial transition.
上皮-间质转化（EMT）是一个重要的生理过程，它决定了肺组织损伤后愈合的结果。诱导EMT的刺激因素和分子级联反应导致肺泡上皮细胞中间质特异性基因的上调以及上皮标志物编码基因的下调。肺泡上皮细胞系通常被用作体外模型来研究肺组织中发生的过程。本研究的目的是量化并比较多种肺上皮细胞系中上皮和间质标志物的mRNA表达水平。
培养肺上皮细胞系L2、R3/1和RLE-6TN。进行重复的mRNA分离、逆转录以及使用针对上皮标志物（E-钙黏蛋白、闭合蛋白和ZO-2）和间质标志物（α-平滑肌肌动蛋白、Ⅲ型胶原和波形蛋白）的引物进行定量PCR。
首先，我们的研究显示，与L2和R3/1细胞相比，RLE-6TN细胞系中的上皮转录本水平更高。其次，我们发现在所有研究的细胞系中，上皮标志物（E-钙黏蛋白、闭合蛋白和ZO-2）和间质标志物（α-平滑肌肌动蛋白、Ⅲ型胶原和波形蛋白）的mRNA同时表达。
我们的数据表明，在转录水平上，L2、R3/1和RLE-6TN细胞系处于EMT的中间阶段之一，这为在细胞系上研究EMT开辟了新的可能性。确定上皮和间质标志物变化的方向将有助于确定导致EMT和反向间质-上皮转化的因素。
