GENE REGULATION IN TERATOCARCINOMA AND EMBRYONIC CELLS

畸胎瘤和胚胎细胞中的基因调控

• 批准号: 3314895
• 负责人: SIDNEY STRICKLAND
• 金额: $ 17.9万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-01-01 至 1990-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3314895
• 关键词: RNA biosynthesis; RNA splicing; autoradiography; cell differentiation; chromatin; collagen; developmental genetics; early embryonic stage; electron microscopy; embryo /fetus cell /tissue; enzyme feedback; gene expression; genetic library; genetic manipulation; genetic regulation; granulosa cell; immunochemistry; indomethacin; laboratory mouse; mammalian embryology; messenger RNA; molecular cloning; neoplasm /cancer genetics; nucleic acid sequence; oligonucleotides; ovary; plasminogen activator; protein biosynthesis; protein engineering; protein structure; proteoglycan; radiotracer; teratoma; tissue /cell culture

Plasminogen activators are serine proteases that have been implicated in a wide variety of normal and abnormal processes. The experiments proposed address two broad questions concerning these enzymes: how is the synthesis of tissue plasminogen activator (tPA) controlled, and what are the respective roles for tPA and the urokinase-type plasminogen activator (uPA). Regulation of tPA synthesis will be studied in F9 teratocarcinoma cells, in which enzyme production can be conveniently controlled by manipulation of the culture conditions. The fact that tPA can feedback inhibit its own synthesis will also be examined in these cells by classical techniques and oligonucleotide-mediated mutagenesis followed by transfection and expression of specifically modified enzymes. The tumorigenic properties of F9 cells producing these modified tPAs will also be determined. In the rat ovary, granulosa cells produce tPA and thecal cells produce uPA. With both cells, enzyme synthesis is maximal at the time of ovulation and is induced by gonadotropins. This system will be used in vivo and in vitro to investigate the function of the two enzymes in the ovulatory process. The possibility that tPA is instrumental in degrading follicular proteoglycans, and that proteoglycans can stimulate the activity of tPA, will be examined.

纤溶酶原激活物是丝氨酸蛋白酶，与一种 各种正常和不正常的过程。建议进行的实验 回答关于这些酶的两个广泛的问题：如何合成 组织纤溶酶原激活物(TPA)的控制，以及 TPA和尿激酶型纤溶酶原激活物(UPA)各自的作用。 将在F9畸胎癌细胞中研究tPA合成的调节。 哪种酶的产生可以通过操纵 培养条件。TPA可以反馈的事实抑制了它自己 合成也将在这些细胞中通过经典技术和 寡核苷酸介导的诱变后的转基因和 表达特定修饰的酶。肿瘤的致癌特性 产生这些修饰的TPA的F9细胞也将被确定。 在大鼠卵巢中，颗粒细胞产生tPA，膜细胞产生 UPA。在这两种细胞中，酶的合成在排卵时达到最大。 并由促性腺激素诱导。该系统将在体内和在 体外研究这两种酶在排卵中的作用 进程。组织型纤溶酶原激活剂在卵泡降解中的作用 蛋白多糖，蛋白多糖可以刺激tPA的活性， 将会被检查。