Transcriptional profiling of mucociliary differentiation in human airway epithelial cells
Transcriptional profiling of mucociliary differentiation in human airway epithelial cells
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DOI：
10.1165/rcmb.2006-0466oc
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发表时间：
2007-08-01
影响因子：
6.4
通讯作者：
Devlin, Robert B.
中科院分区：
文献类型：
--
作者：
Ross, Andrea J.;Dailey, Lisa A.;Devlin, Robert B.
当在适当的培养基中的气液界面（ALI）中培养时，原代人气道上皮细胞形成由纤毛细胞和粘液分泌细胞组成的极化、假复层上皮。该培养系统为气道上皮生物学和分化的体外研究提供了有用的工具。我们对生长了 28 天的人支气管上皮细胞 (HBEC) 的 ALI 培养物进行了微阵列分析，以鉴定参与粘膜纤毛分化的基因。我们鉴定了超过 2,000 个基因，这些基因在一段时间内表现出统计上显着的 2 倍或更大的表达变化。在显示出最大增加的基因中，许多基因涉及与气道上皮生物学相关的过程，例如细胞粘附、免疫、运输和纤毛形成；然而，许多新基因也被鉴定出来。我们将我们的结果与纤毛轴丝蛋白质组学分析的数据进行比较，并确定了可能在纤毛形成或功能中发挥作用的候选基因。使用 Ingenuity Pathways Analysis（Ingenuity Systems，Redwood City，CA）生成基因网络，以识别参与粘液纤毛细胞分化或功能的信号传导途径。鉴定出包含 TGF-β、WNT/β-连环蛋白和表皮生长因子受体 (EGFR) 通路相关基因的网络，表明这些家族在气道上皮细胞中的潜在作用。通过实时 RT-PCR 对一些代表性基因的微阵列结果进行了验证。这项工作提供了有关气道上皮细胞分化过程中基因表达变化的广泛信息，对于对呼吸功能、病理学和毒理学感兴趣的研究人员来说将是有用的资源。
When cultured at an air-liquid interface (ALI) in the appropriate medium, primary human airway epithelia[ cells form a polarized, pseudostratified epithelium composed of ciliated and mucus-secreting cells. This culture system provides a useful tool for the in vitro study of airway epithelial biology and differentiation. We have performed microarray analysis on ALI cultures of human bronchial epithelial cells (HBECs) grown over a 28-d period to identify genes involved in mucociliary differentiation. We identified over 2,000 genes that displayed statistically significant 2-fold or greater changes in expression during the time course. Of the genes showing the largest increases, many are involved in processes associated with airway epithelial biology, such as cell adhesion, immunity, transport, and cilia formation; however, many novel genes were also identified. We compared our results with data from proteomic analyses of the ciliary axoneme and identified candidate genes that may have roles in cilia formation or function. Gene networks were generated using Ingenuity Pathways Analysis (Ingenuity Systems, Redwood City, CA) to identify signaling pathways involved in mucociliary cell differentiation or function. Networks containing genes involved in TGF-beta, WNT/beta-catenin, and epidermal growth factor receptor (EGFR) pathways were identified, suggesting potential roles for these families in airway epithelia. Microarray results were validated by real-time RT-PCR for a number of representative genes. This work has provided extensive information about gene expression changes during differentiation of airway epithelial cells, and will be a useful resource for researchers interested in respiratory function, pathology, and toxicology.