Proteomics-based identification of proteins secreted in apical surface fluid of squamous metaplastic human tracheobronchial epithelial cells cultured by three-dimensional organotypic air-liquid interface method
Proteomics-based identification of proteins secreted in apical surface fluid of squamous metaplastic human tracheobronchial epithelial cells cultured by three-dimensional organotypic air-liquid interface method
复制标题
DOI：
10.1158/0008-5472.can-06-2783
复制
发表时间：
2007-07-15
期刊：
影响因子：
11.2
通讯作者：
Koo, Ja Seok
中科院分区：
文献类型：
--
作者：
Kim, Seung-Wook;Cheon, Kyounga;Koo, Ja Seok
肺鳞状细胞癌起源于支气管上皮细胞，这些细胞获得越来越多的异常表型。目前，没有已知的生物标志物是临床有效的癌前病变和肺癌的早期检测。我们试图确定分泌的分子产生的鳞状支气管上皮细胞培养的器官型培养方法。我们分析了异常分化鳞状化生正常人气管支气管上皮（NHTBE）和粘液NHTBE细胞的顶面液（ASF）中的蛋白表达模式。比较二维PAGE分析显示，174个独特的蛋白质在ASF的鳞状NHTBE细胞与正常粘膜纤毛分化的NHTBE细胞相比。其中，64个分离良好的蛋白质点，通过液相色谱-串联质谱鉴定，揭示了22个不同的蛋白质从鳞状NHTBE细胞的ASF。用免疫印迹分析进一步证实了鳞状NHTBE细胞中这些蛋白质中的六种[SCC抗原1（SCCA 1）、SCC抗原2（SCCA 2）、S100 A8、S100 A9、膜联蛋白I和膜联蛋白II]的表达。值得注意。SCCA 1和SCCA 2被证实在鳞状化生的NHTBE细胞中表达，但在正常粘膜NHTBE或正常支气管上皮中不表达。此外，SCCA 1和SCCA 2的表达在体外肺癌模型细胞系中随着恶性程度的增加而增加。总之，我们鉴定了通过器官型气液界面方法培养的鳞状化生原代人支气管上皮细胞独特分泌的蛋白质。这些ASF蛋白可用于检测肺部异常病变，而无需采集侵入性活检标本。
Squamous cell carcinoma in the lung originates from bronchial epithelial cells that acquire increasingly abnormal phenotypes. Currently, no known biomarkers are clinically efficient for the early detection of premalignant lesions and lung cancer. We sought to identify secreted molecules produced from squamous bronchial epithelial cells cultured with organotypic culture methods. we analyzed protein expression patterns in the apical surface fluid (ASF) from aberrantly differentiated squamous metaplastic normal human tracheobronchial epithelial (NHTBE) and mucous NHTBE cells. Comparative two-dimensional PAGE analysis revealed 174 unique proteins in the ASF of squamous NHTBE cells compared with normal mucociliary differentiated NHTBE cells. Among them, 64 well-separated protein spots were identified by liquid chromatography-tandem mass spectrometry, revealing 22 different proteins in the ASF from squamous NHTBE cells. Expression of six of these proteins [SCC antigen 1 (SCCA1), SCC antigen 2 (SCCA2), S100A8, S100A9, Annexin I, and Annexin II] in the squamous NHTBE cells was further confirmed with immunoblot analysis. Notablv. SCCA1 and SCCA2,were verified as being expressed in squamous metaplastic NHTBE cells but not in normal mucous NHTBE or normal bronchial epithelium. Moreover, SCCA1 and SCCA2 expression increased in in vitro lung carcinogenesis model cell lines with increasing malignancy. In summary, we identified proteins that are uniquely secreted from squamous metaplastic primary human bronchial epithelial cells cultured by the organotypic air-liquid interface method. These ASF Proteins may be used to detect abnormal lesions in the lung without collecting invasive biopsy specimens.