DIFFERENTIATION AND DIFFERENTIATIVE FUNCTIONS OF AIRWAY EPITHELIAL CELLS

气道上皮细胞的分化和分化功能

• 批准号: 3965231
• 负责人: A 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/3965231
• 关键词: affinity chromatography; antibacterial agents; cancer prevention; carcinogenesis inhibitor; cell differentiation; cell population study; gel electrophoresis; gene expression; hyaluronate; keratinization; molecular weight; monoclonal antibody; mucins; neoplasm /cancer nutrition therapy; neoplastic transformation; nutrition related tag; oligosaccharides; protein glutamine gamma glutamyltransferase; radiotracer; respiratory epithelium; secretion; sialate; squamous cell carcinoma; tissue /cell culture; trachea; trachea neoplasm; tumor promoters; vitamin therapy

Rabbit tracheal epithelial cells grown on fibronectin-albumin-vitrogen coated dishes in serum-free and vitamin A-free medium undergo terminal squamous differentiation under a variety of conditions. This differentiation occurs in several stages. First cells undergo terminal cell division; cells accumulate in G0/G1 phase of the cell cycle and loose the ability to form colonies. This terminal cell division occurs at high density or is induced at low density when epidermal growth factor is omitted from the medium or when TGFBeta is added. This is followed by the expression of a squamous phenotype that is characterized by the induction of a squamous morphology, increase in transglutaminase and cholesterol sulfate levels, increase in the expression of a 48 and 56 kd keratin and the formation of cross-linked envelopes. The increase in cholesterol sulfate levels appears to be related to an enhancement in sulfotransferase activity. High calcium concentration promotes the expression of the squamous phenotype. Retinoic acid has no effect on the commitment to terminal cell division but inhibits the expression of the squamous phenotype; retinoic acid inhibits the increase in transglutaminase and cholesterol sulfate levels the alterations in keratin expression and the formation of cross-linked envelopes. To study the regulation of differentiation at the molecular level, we established a cDNA library using poly (A)+ RNA from squamous differentiated cells and isolated recombinant cDNA clones that hybridize with mRNA's isolated from squamous differentiated cells but not from undifferentiated and retinoic acid-treated cells. This indicates that these clones represent gene products that function as markers for differentiation. We are in the process to analyze at what level these gene products are regulated and how retinoids control them.

在纤连蛋白-白蛋白-vitrogen 上生长的兔气管上皮细胞 在无血清和无维生素 A 的培养基中进行涂层培养皿的最终处理 不同条件下的鳞状细胞分化。 这 分化发生在几个阶段。 第一个细胞经历终末期 细胞分裂；细胞在细胞周期的G0/G1期积累并释放 形成殖民地的能力。 这种终末细胞分裂发生在高 密度或当表皮生长因子在低密度下诱导 从培养基中省略或添加 TGFBeta 时。 接下来是 鳞状表型的表达，其特征在于诱导 鳞状形态，转谷氨酰胺酶和胆固醇增加 硫酸盐水平，48 和 56 kd 角蛋白表达增加， 交联包膜的形成。 胆固醇增加 硫酸盐水平似乎与磺基转移酶的增强有关 活动。 高钙浓度促进表达 鳞状表型。 视黄酸对承诺没有影响 终末细胞分裂但抑制鳞状细胞的表达 表型；视黄酸抑制转谷氨酰胺酶的增加 硫酸胆固醇水平改变角蛋白表达和 交联包膜的形成。 研究监管 为了在分子水平上进行分化，我们使用以下方法建立了 cDNA 文库： 来自鳞状分化细胞的聚 (A)+ RNA 和分离的重组体 与从鳞状细胞中分离的 mRNA 杂交的 cDNA 克隆 分化细胞，但不是未分化细胞和视黄酸细胞 酸处理的细胞。 这表明这些克隆代表基因 作为差异化标志的产品。 我们在 分析这些基因产物在什么水平上受到调节以及如何调节的过程 类维生素A控制它们。