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AN ADRENAL TUMOR: CYTOCHROME P-450 AND STEROIDOGENESIS

肾上腺肿瘤：细胞色素 P-450 和类固醇生成

基本信息

批准号：
3168724
负责人：
PETER F HALL
金额：
$ 15.66万
依托单位：
WORCESTER FOUNDATION FOR BIOMEDICAL RES
依托单位国家：
美国
项目类别：
财政年份：
1980
资助国家：
美国
起止时间：
1980-07-01 至 1986-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3168724
关键词：
adrenal neoplasm adrenocorticotropic hormone aminoacid analyzer cholera toxin cholesterol cytochrome P450 hormone regulation /control mechanism host neoplasm interaction hydroxysteroids membrane activity mitochondria neoplasm /cancer immunology radioimmunoassay steroid biosynthesis steroid hormone biosynthesis tissue /cell culture

项目摘要

Mouse adrenal tumor cells (Y-1) in monolayer culture is being used to study questions related to their invasive properties, to the regulation of their steroidogenic activity, and to the relationship between these two features of the cells as follows: What is the function of the newly synthesized proteins found associated with mitochondria after addition of ACTH to Y-1 cells? What effect do alterations in plasma membrane viscosity and relocation of membrane receptors in an electric field have on the initial events associated with responses of the cells to ACTH and cholera toxin? To what extent can the enhancement of growth and steroidogenesis produced by injection of cells into LAF1 mice be duplicated by addition of pituitary extracts to the cells in culture? The relationship between the new protein(s) and transport of cholesterol to mitochondria is studied by incubating [3H]cholesterol oleate--low density lipoprotein with cells in which side-chain cleavage is blocked and measuring mitochondrial [3H]cholesterol. Membrane receptors are labeled with fluorescent ligands (ACTH, cholera toxin, and Concanavalin A) after exposure to an electrical field to estimate membrane viscosity. Cholesterol content of cell membranes is altered by incubation with lipid-free plasma or cholesterol-containing liposomes. Steroidogenesis and growth of cells in the presence of pituitary extracts are compared with the response to animal passage. Nonionic detergents are used to improve extraction of mitochondrial P-450 from Y-1 cells and normal adrenal cells. The role of myosin in the response to ACTH is studied by measuring the extent of phosphorylation of the myosin light chain when cells are incubated without and with ACTH. A cyclic AMP-dependent protein kinase has been found closely associated with the cytoskeleton of Y-1 cells. The enzyme promotes phosphorylation of seven proteins. Protein kinase C has also been found in Y-1 cells which show increased steroidogenesis when incubated with phorbol esters. (D)

单层培养的小鼠肾上腺肿瘤细胞（Y-1）被用于研究有关其侵入性的问题，对他们的监管，类固醇生成活性，以及这两个特征之间的关系细胞的功能如下：新合成的在Y-1中加入ACTH后发现与线粒体相关的蛋白质细胞？什么样的影响改变质膜粘度和膜受体在电场中的重新定位对初始的与细胞对ACTH和霍乱毒素的反应有关的事件？在何种程度上可以促进生长和类固醇生成通过将细胞注入LAF1 小鼠通过添加垂体提取物对培养细胞的作用新的蛋白质与胆固醇转运到线粒体的研究是通过孵育[3胆固醇油酸酯--低密度脂蛋白与其中侧链裂解被阻断的细胞，测量线粒体[3H]胆固醇。膜受体被标记与荧光配体（ACTH，霍乱毒素和伴刀豆球蛋白A），暴露于电场以估计膜粘度。细胞膜的胆固醇含量通过与以下物质孵育而改变：无脂质血浆或含胆固醇的脂质体。类固醇生成和细胞生长的垂体提取物的存在下进行了比较，对动物传代的反应。非离子洗涤剂用于改善从Y-1细胞和正常肾上腺细胞中提取线粒体P-450。肌球蛋白在对ACTH的反应中的作用是通过测量肌球蛋白轻链的磷酸化程度，不与ACTH一起孵育和与ACTH一起孵育。环腺苷酸依赖性蛋白激酶已被发现密切相关 Y-1细胞的细胞骨架该酶促进磷酸化，七种蛋白质在Y-1细胞中也发现了蛋白激酶C，当与佛波酯一起孵育时，显示类固醇生成增加。（丁）