KERATINOCYTES--MODULATION BY RETINOIC ACID

角质细胞——视黄酸的调节

• 批准号: 3457306
• 负责人: Noreen J Hickok
• 金额: $ 11.45万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3457306
• 关键词: DNA footprinting; acne; aging; basal cell carcinoma; binding proteins; chloramphenicol; complementary DNA; congenital ichthyosis; cycloheximide; decarboxylase inhibitor; estrogen receptors; fibronectins; gel mobility shift assay; gene induction /repression; growth inhibitors; keratinocyte; light adverse effect; messenger RNA; nutrition related tag; ornithine decarboxylase; plasmids; protein glutamine gamma glutamyltransferase; psoriasis; retinoate; skin disorder; skin disorder chemotherapy; thymidine kinase; transcription factor; vitamin therapy

Retinoids are a class of compounds which are potent therapeutic agents for a number of dermatological diseases. In order to improve treatment of these diseases, it is necessary to elucidate the mechanism whereby retinoids, such as all-trans-retinoic acid (RA), exert their anti- proliferative effects. The long term goal of these studies is, therefore, to determine how RA inhibits keratinocyte growth. An important action of RA is keratinocytes is to inhibit ornithine decarboxylase (OD), an enzyme whose products, the polyamines, are necessary for cell growth. It is therefore proposed to determine if inhibition of ODC gene expression is a direct consequence of retinoid action and how this repression is exerted. These experiments re designed to test the hypothesis that RA suppresses ODC gene transcription through a RA receptor (RAR)-mediated mechanism. This will be examined by (1) determining the effects of RA and/or cycloheximide on the relative transcription rates of the human ODC gene using run-on transcription; (2) determining which RAR is expressed in keratinocytes and discriminating between the actions of the cellular RA binding protein and the alpha-, beta-, and gamma-RARs through the use of chimeric RAR/estrogen receptors that will be transfected into keratinocytes; (3) identifying cis- acting DNA elements that interact with trans-acting factors to cause RA- mediated changes in ODC gene transcription, using in vitro footprinting, gel retardation assays, and transfectin of isolated elements ligated to the thymidine kinase-beta-globin gene; and (4) isolation of cDNAs encoding non- RAR trans-acting factors which are implicated in RA action. Elucidation of the mechanisms whereby RA suppresses ODC gene expression will yield important information on how retinoids exert their anti-proliferative effects.

性类视感是一类化合物，它们是有效的治疗剂 许多皮肤病。 为了改善治疗 这些疾病，有必要阐明 类视网膜类似，例如全反击酸（RA），抗抗 增殖作用。 因此，这些研究的长期目标是 确定RA如何抑制角质形成细胞的生长。 一个重要的动作 RA是角质形成细胞是抑制鸟类脱羧酶（OD），一种酶 其产品，多胺是细胞生长所必需的。 这是 因此，建议确定ODC基因表达的抑制是否为A 类视网膜类似作用的直接结果以及如何施加这种抑制作用。 这些实验旨在测试RA抑制ODC的假设 通过RA受体（RAR）介导的机制转录基因转录。 这 将通过（1）确定RA和/或环己酰亚胺的影响来检查 使用跑步的人类ODC基因的相对转录速率 转录； （2）确定在角质形成细胞中表达的RAR和 区分细胞RA结合蛋白的作用和 通过使用嵌合RAR/雌激素的α-，β-和伽马射线 将转染成角质形成细胞的受体； （3）确定顺式 与反式因子相互作用的作用DNA元件，引起ra- 使用体外足迹介导的ODC基因转录变化， 凝胶延迟测定和连接到该元素的孤立蛋白 胸苷激酶 - β-珠蛋白基因； （4）隔离编码非 - 的cDNA 涉及RA作用的RAR反式作用因子。 阐明 RA抑制ODC基因表达的机制将产生 关于类视黄素如何发挥其抗增殖性的重要信息 效果。