Regulation Of Differentiation In Lung Keratinocytes

肺角质形成细胞分化的调节

• 批准号: 7007108
• 负责人: Anton M Jetten
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7007108
• 关键词: cell differentiation; cell growth regulation; esophagus; gene expression; genetic regulation; genetically modified animals; human tissue; keratinocyte; laboratory mouse; lung; respiratory epithelium; respiratory function; retinoids; skin; tissue /cell culture; trachea; transcription factor

I. The trachea and esophagus have a common origin evolving from the foregut endoderm during early embryonic development. These epithelia undergo a series of well-defined structural changes involving differentiation of progenitor cells into several cell types that ultimately result in the formation of the mature epithelium. In this study, we monitor the expression of p63 in the esophageal and tracheobronchial epithelium at several stages during development and determine the effect of the lack of p63 expression on the morphogenesis of these epithelia in p63-/- mice. At day E15.5, the esophageal and tracheobronchial epithelium contain 2-3 layers of cells; however, only the progenitor cells express p63. The progenitor cells differentiate first into ciliated cells (p63-/b-tubulin IV+) and at birth into basal cells (p63+/K14+/BS-I-B4+). In adult mouse and human, the lining of the esophagus matures into a (non)keratinizing, stratified epithelium while the tracheobronchial lining develops into a pseudostratified, columnar epithelium containing basal, ciliated and mucosecretory cells. In mature epithelia, the K14+/BS-I-B4+ basal cells are the most intensely stained for p63. Expression of p63 is dramatically repressed during squamous differentiation in vivo or in cultured cells. Generally, human squamous cell carcinomas stained strongly while human adenocarcinomas did not stain for p63. In contrast to the esophagus and trachea from wild type mice, the esophageal and tracheobronchial epithelium from newborn p63-/- mice consist largely of a columnar, ciliated epithelium that appear to lack basal cells. Our study indicates that p63 is critical for normal morphogenesis of the esophageal and tracheobronchial epithelium. In p63-/- mice, progenitor cells are able to differentiate into ciliated cells but do not appear to generate basal cells suggesting a role for p63 either in the regulation of the differentiation of progenitor cells into basal cells or in the survival of basal cells. II. Retinoids play an important role in the tracheobronchial epithelium. Retinoids can act via nuclear retinoid receptors or through other mechanisms. The synthetic retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (AHPN) can act both activate RARgamma and act by receptor-independent mechanisms. We have analyzed the effect of several retinoids on the expression of non-steroidal anti-inflammatory drug-activated gene (NAG-1) in normal human tracheobronchial epithelial (HTBE) cells and several lung carcinoma cell lines. The retinoid AHPN greatly enhances the expression of NAG-1 mRNA and protein in a time and dose-dependent manner in human lung adenocarcinoma H460 cells and several other carcinoma cell lines. This induction was specific for AHPN since retinoic acid, an RAR- and an RXR-pan-agonist were unable to induce NAG-1 suggesting that this induction is not mediated through activation of retinoid receptors. Although NAG-1 is a p53-responsive gene, AHPN-induced NAG-1 expression does not require p53. The induction of NAG-1 expression by AHPN is at least in part due to a 8-fold increase in the stability of NAG-1 mRNA. In contrast to carcinoma cells, NAG-1 expression is effectively induced by retinoic acid and the RAR-selective pan-agonist in normal HTBE cells and accompanies the inhibition of squamous differentiation and the initiation of normal differentiation. In vivo, NAG-1 expression was observed in the normal tracheobronchial epithelium while no expression was found in either squamous metaplastic tracheal epithelium or in sections of human lung tumors. Our results suggest that the induction of NAG-1 expression by retinoids in normal HTBE and lung carcinoma cells is regulated by distinct mechanisms and is associated with different biological processes. The linkage between AHPN treatment and NAG-1 expression revealed in this study provides a new mechanism for the anti-tumorigenic activity of AHPN.

I.气管和食管具有共同的起源，在早期胚胎发育期间从前肠内胚层进化而来。这些上皮细胞经历一系列明确的结构变化，包括祖细胞分化成几种细胞类型，最终导致成熟上皮的形成。在这项研究中，我们监测p63在食管和气管支气管上皮细胞在几个阶段的发展过程中的表达，并确定这些上皮细胞的形态发生在p63-/-小鼠的p63表达的缺乏的影响。在E15.5天，食管和气管支气管上皮含有2-3层细胞;然而，只有祖细胞表达p63。祖细胞首先分化成纤毛细胞（p63-/b-微管蛋白IV+），并在出生时分化成基底细胞（p63+/K14+/BS-I-B4+）。在成年小鼠和人中，食管衬里成熟为（非）角化、复层上皮，而气管支气管衬里发育为假复层、柱状上皮，含有基底、纤毛和粘膜分泌细胞。在成熟上皮中，K14+/BS-I-B4+基底细胞对p63的染色最强。在体内或培养的细胞中，p63的表达在鳞状分化期间被显著抑制。一般来说，人鳞状细胞癌染色强烈，而人腺癌不染色p63。与野生型小鼠的食管和气管相反，新生p63-/-小鼠的食管和气管支气管上皮主要由柱状纤毛上皮组成，似乎缺乏基底细胞。我们的研究表明，p63是食管和气管支气管上皮细胞的正常形态发生的关键。在p63-/-小鼠中，祖细胞能够分化成纤毛细胞，但似乎不产生基底细胞，这表明p63在调节祖细胞分化成基底细胞或基底细胞存活中的作用。 二.维甲酸在气管支气管上皮中起重要作用。类维生素A可以通过核类维生素A受体或通过其他机制发挥作用。合成的类维生素A 6-[3-（1-金刚烷基）-4-羟基苯基]-2-萘羧酸（AHPN）可以激活RAR γ并通过受体非依赖性机制起作用。我们分析了几种维甲酸对正常人气管支气管上皮细胞（HTBE）和几种肺癌细胞系中非甾体抗炎药激活基因（NAG-1）表达的影响。维甲酸AHPN可显著增强人肺腺癌H460细胞及其他几种肿瘤细胞株NAG-1 mRNA和蛋白的表达，并呈时间和剂量依赖性。这种诱导对AHPN是特异性的，因为视黄酸、RAR和RXR泛激动剂不能诱导NAG-1，表明这种诱导不是通过类视色素受体的激活介导的。虽然NAG-1是一个p53应答基因，但AHPN诱导的NAG-1表达并不需要p53。AHPN诱导NAG-1表达至少部分是由于NAG-1 mRNA稳定性增加8倍。与癌细胞相反，NAG-1表达在正常HTBE细胞中被视黄酸和RAR选择性泛激动剂有效诱导，并伴随鳞状分化的抑制和正常分化的启动。在体内，NAG-1的表达被观察到在正常气管支气管上皮细胞，而没有表达被发现在鳞状化生气管上皮细胞或在人类肺肿瘤的部分。我们的研究结果表明，在正常HTBE和肺癌细胞中，类维生素A对NAG-1表达的诱导受不同机制的调节，并与不同的生物学过程相关。本研究揭示的AHPN治疗与NAG-1表达之间的联系为AHPN的抗肿瘤活性提供了新的机制。

项目成果

Induction of the cytochrome P450 gene CYP26 during mucous cell differentiation of normal human tracheobronchial epithelial cells.

正常人气管支气管上皮细胞粘液细胞分化过程中细胞色素 P450 基因 CYP26 的诱导。

• DOI: 10.1124/mol.58.3.483
• 发表时间: 2000
• 期刊: Molecular pharmacology
• 影响因子: 3.6
• 作者: Kim,SY;Adachi,H;Koo,JS;Jetten,AM
• 通讯作者: Jetten,AM