MOLECULAR GENETIC STUDIES OF THYROID CELL LINES

这些研究的长期目标是建立自发的 猫功能亢进性腺瘤性甲状腺肿(毒性结节性甲状腺肿) 人甲状腺良性肿瘤模型的建立及分子遗传学研究 与从正常到腺瘤转换相关的异常 保持分化功能的生长。建议进行的研究 关注控制猫科动物正常细胞周期的因素 腺瘤性甲状腺细胞系。我们将特别注意 癌基因和抑癌基因结构可能的改变和 表情。为了获得必要的技术来完成 这一长期目标，这项提案概述了试点工作，将允许 结合分子遗传学技术的首席研究人员 正在进行的和未来的甲状腺学研究项目。 在涉及猫腺瘤甲状腺细胞系的实验中 PETCAT-L、-2、-3、-4和ROMCAT-1，该提案具有以下具体内容 目标： 1.让来访的研究人员熟悉分子 核糖核酸和脱氧核糖核酸提取纯化的基因技术 酸性电泳，斑点印迹分析，Northern和Southern杂交 杂交技术，并学习分子生物学的应用 基因技术用于研究细胞周期事件。 2.检测大鼠肝细胞DNA结构及基础mRNA表达的变化。 已知的原癌基因、转录因子、肿瘤抑制因子、 和信号转导因子。 3.研究生长相关遗传因子的基因表达动力学 并将其与甲状腺细胞相关因子的表达进行比较 在促进生长的条件下分化。

The longterm objectives of these studies are to establish spontaneous feline hyperfunctional adenomatous goiter (toxic nodular goiter) as a model for human benign thyroid tumors, and to probe for molecular genetic abnormalities associated with the changeover from normal to adenomatous growth with maintained differentiated function. The proposed studies focus upon the factors controlling the normal cell cycle of the feline adenomatous thyrocyte cell lines. Particular attention will be paid to possible alterations in oncogene and anti-oncogene structure and expression. In order to acquire the necessary techniques to accomplish this longterm goal, this proposal outlines pilot work which will allow the principal investigator to incorporate molecular genetic techniques into ongoing and future research projects in thyroidology. In experiments involving the feline adenomatous thyrocyte cell lines PETCAT-l,-2,-3,-4, and ROMCAT-1, this proposal has the following specific aims: 1. For the visiting researcher to become familiar with the molecular genetic techniques of RNA and DNA extraction and purification, nucleic acid electrophoresis, dot blot assays, Northern and Southern blotting and hybridization techniques, and to learn about application of molecular genetic techniques to the study of cell cycle events. 2. To evaluate alterations in DNA structure and basal mRNA expression of known proto-oncogenes, transcription factors, tumor suppression factors, and signal transduction factors. 3. To study mRNA expression kinetics of growth-related genetic factors and compare them with the expression of factors associated with thyrocyte differentiation under conditions stimulatory for growth.