Restoration of the mucous phenotype by retinoic acid in retinoid-deficient human bronchial cell cultures: Changes in mucin gene expression
Restoration of the mucous phenotype by retinoic acid in retinoid-deficient human bronchial cell cultures: Changes in mucin gene expression
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DOI：
10.1165/ajrcmb.20.1.3310
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发表时间：
1999-01-01
影响因子：
6.4
通讯作者：
Nettesheim, P
中科院分区：
文献类型：
--
作者：
Koo, JS;Yoon, JH;Nettesheim, P
类维生素A缺乏的气道上皮细胞培养物经历鳞状分化。用视黄酸（RA）处理这些培养物导致粘液表型的恢复。本研究的目的是描述类维生素A缺乏的人气管支气管上皮细胞培养物经RA治疗后的细胞和分子变化。特别感兴趣的是确定在鳞状细胞向粘液细胞表型转化期间，粘蛋白基因MUC 2、MUC 5AC和MUC 5 B何时表达。我们使用角蛋白α和分泌粘蛋白作为标记物来监测鳞状和粘液表型，分别。我们的研究表明，RA反应的文化进行性下降与长期类维生素A缺乏症，需要更高的RA浓度，以恢复粘液表型。RA治疗开始后12小时内，角蛋白ct表达下降，表明细胞表型开始发生变化。在24小时后，RA的培养物中，出现了大量的粘液细胞，并在72小时粘蛋白分泌可测量的量。粘蛋白基因表达的诱导依次发生：MUC 2、MUC 5AC和MUC 5 B mRNA分别在24、48和72 h上调。当用10（-6）M RA处理10（-8）M RA中的培养物时，MUC 2而不是MUC 5AC和MUC 5 B mRNA水平在6小时内上调。我们的研究表明，MUC 2 mRNA是粘液分化的早期标志物，而MUC 5AC和MUC 5 B mRNA在粘液分化的更高级阶段表达。我们的研究进一步表明，每个粘蛋白基因是由不同的机制。
Retinoid-deficient cultures of airway epithelial cells undergo squamous differentiation. Treatment of such cultures with retinoic acid (RA) leads to restoration of the mucous phenotype. The purpose of our study was to characterize the cellular and molecular changes following RA treatment of retinoid-deficient human tracheobronchial epithelial cell cultures. Of particular interest was to determine when during the conversion of the squamous to the mucous phenotype the mucin genes MUC2, MUC5AC, and MUC5B were expressed. We used cornifin alpha and secreted mucin as markers to monitor the squamous and mucous phenotypes, respectively. Our studies showed that the RA responsiveness of the cultures progressively decreased with protracted retinoid deficiency, requiring higher RA concentrations to restore the mucous phenotype. Within 12 h after the start of RA treatment, cornifin ct expression decreased, signaling the beginning of a change in cellular phenotype. At 24 h after addition of RA to the cultures, a significant number of mucous cells appeared, and at 72 h mucin was secreted in measurable amounts. Induction of mucin gene expression occurred sequentially: MUC2, MUC5AC, and MUC5B mRNAs were upregulated at 24, 48, and 72 h, respectively. When cultures maintained in 10(-8) M RA were treated with 10(-6) M RA, MUC2 but not MUC5AC and MUC5B mRNA levels were upregulated within 6 h. Our study indicates that MUC2 mRNA is an early marker of mucous differentiation, whereas MUC5AC and MUC5B mRNAs are expressed during more advanced stages Of mucous differentiation. Our studies further suggest that each of the mucin genes is regulated by distinct mechanisms.