Polosa R, Prosperini G, Tomaselli V, Howarth P H, Holgate S T, Davies D E
Division of Respiratory, Cell & Molecular Biology, Faculty of Medicine, University of Southampton, Southampton General Hospital, Southampton, United Kingdom.
J Allergy Clin Immunol. 2000 Dec;106(6):1124-31. doi: 10.1067/mai.2000.110931.
The epidermal growth factor (EGF) family of growth factors plays an important role in maintenance and repair in a variety of epithelial tissues. However, very little is known about coexpression of these factors and their receptors, the c-erbB family of receptor tyrosine kinases, in human nasal epithelium.
We sought to investigate the expression of these molecules in cultured nasal epithelial cells and nasal mucosa from healthy individuals.
Identification of c-erbB receptors and their ligands was sought by using reverse transcription PCR, Western blotting, and immunohistochemistry.
Messenger RNA encoding the EGF receptors (EGFR) c-erbB2 and c-erbB3, but not c-erbB4, was detected in primary cultures of human nasal epithelial cells. Transcripts encoding EGF, heparin-binding EGF, transforming growth factor (TGF) alpha, and amphiregulin were also detected. Receptor and ligand expression was confirmed by using immunocytochemical staining of the cells and Western blotting of the cell lysates. Immunohistochemical analysis of tissue sections obtained from biopsy specimens of nasal mucosa revealed intense membrane staining for the EGFR within the respiratory nasal epithelium, which was predominantly localized at the level of the columnar epithelial layers. Similar staining patterns were observed for c-erbB2 and c-erbB3 in the respiratory nasal epithelium. Evidence for EGF, transforming growth factor alpha, heparin-binding EGF, amphiregulin, and betacellulin immunostaining in the nasal epithelium was also obtained; their staining patterns paralleled that of EGFR immunostaining.
Colocalization of c-erbB receptors and ligands establishes a basis on which to investigate c-erbB receptor- mediated effects in human nasal epithelium.
表皮生长因子（EGF）家族在多种上皮组织的维持和修复中发挥重要作用。然而，对于这些因子及其受体（受体酪氨酸激酶的c-erbB家族）在人鼻上皮中的共表达情况，人们了解甚少。
我们试图研究这些分子在健康个体培养的鼻上皮细胞和鼻黏膜中的表达。
通过逆转录聚合酶链反应、蛋白质印迹法和免疫组织化学法来鉴定c-erbB受体及其配体。
在人鼻上皮细胞原代培养物中检测到编码表皮生长因子受体（EGFR）c-erbB2和c-erbB3的信使核糖核酸，但未检测到c-erbB4。还检测到了编码表皮生长因子、肝素结合表皮生长因子、转化生长因子（TGF）α和双调蛋白的转录本。通过对细胞进行免疫细胞化学染色和对细胞裂解物进行蛋白质印迹法，证实了受体和配体的表达。对鼻黏膜活检标本的组织切片进行免疫组织化学分析显示，呼吸性鼻上皮内的EGFR有强烈的膜染色，主要定位于柱状上皮层水平。在呼吸性鼻上皮中，c-erbB2和c-erbB3也观察到类似的染色模式。在鼻上皮中也获得了表皮生长因子、转化生长因子α、肝素结合表皮生长因子、双调蛋白和β细胞素免疫染色的证据；它们的染色模式与EGFR免疫染色的模式相似。
c-erbB受体与配体的共定位为研究c-erbB受体介导的人鼻上皮效应奠定了基础。
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