Koslowski Roland, Barth Kathrin, Augstein Antje, Tschernig Thomas, Bargsten Gerhard, Aufderheide Michaela, Kasper Michael
Institute of Physiological Chemistry, Medical Faculty Carl Gustav Carus, Dresden University of Technology, Fiedlerstrasse 42, 01307 Dresden, Germany.
Histochem Cell Biol. 2004 Jun;121(6):509-19. doi: 10.1007/s00418-004-0662-4. Epub 2004 Jun 8.
The study of function and regulation of the phenotype of alveolar type I (AT I) epithelial cells is limited by the rareness of suitable cell lines or primary cultures of this cell type. We describe in the present study the type I-like rat epithelial cell line R3/1. This cell line displays in vitro a phenotype with several characteristic features of AT I cells. R3/1 cells were analysed for mRNA and protein content of markers related to the AT I cell type (T1alpha, ICAM-1, connexin-43, caveolins-1 and -2) and AT II phenotypes [surfactant proteins (SPs) A, B, C and D]. The mRNAs for SPs were found to be at a low level. Moderate protein levels for SP-A and SP-B were found, and SP-C and SP-D proteins were not detectable. R3/1 cells are positive for CD44s, E-cadherin, cytokeratin, vimentin and RAGE, and bind the lectins BPA and SBA. For demonstration of the suitability of R3/1 cells for in vitro studies on epithelial injury, the cells were treated with bleomycin. As shown by real-time RT-PCR and immunoblotting, bleomycin-treatment of R3/1 cells resulted in a decrease in mRNA and protein for both caveolin-1 and caveolin-2 in comparison with controls. The AT I-like cell line R3/1 may serve as a promising tool for the study of lung cell biology.
肺泡I型(AT I)上皮细胞表型的功能和调控研究受到该细胞类型合适细胞系或原代培养物稀缺性的限制。在本研究中,我们描述了I型样大鼠上皮细胞系R3/1。该细胞系在体外表现出具有AT I细胞若干特征的表型。对R3/1细胞进行了与AT I细胞类型相关标志物(T1α、ICAM-1、连接蛋白-43、小窝蛋白-1和-2)以及AT II表型[表面活性蛋白(SPs)A、B、C和D]的mRNA和蛋白质含量分析。发现SPs的mRNA水平较低。检测到SP-A和SP-B的蛋白水平中等,而未检测到SP-C和SP-D蛋白。R3/1细胞对CD44s、E-钙黏蛋白、细胞角蛋白、波形蛋白和RAGE呈阳性,并能结合凝集素BPA和SBA。为证明R3/1细胞适用于上皮损伤的体外研究,用博来霉素处理这些细胞。实时RT-PCR和免疫印迹结果显示,与对照组相比,博来霉素处理的R3/1细胞中,小窝蛋白-1和小窝蛋白-2的mRNA和蛋白水平均降低。I型样细胞系R3/1可能成为研究肺细胞生物学的一个有前景的工具。