MECHANISMS OF LUNG ADENOMA FORMATION

肺腺瘤形成机制

• 批准号: 3071531
• 负责人: ALVIN M MALKINSON
• 金额: $ 4.96万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-01 至 1989-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3071531
• 关键词: antibody formation; butylated hydroxytoluene; chemical carcinogen; chemical carcinogenesis; cyclic AMP; gene expression; high performance liquid chromatography; laboratory mouse; laboratory rat; lung neoplasms; neoplasm /cancer genetics; polyurethanes; protein kinase A; radiotracer

An RCDA is requested to relieve a heavy teaching load, and allow full-time research on the project described below as well as time to gain experience in recombinant DNA technology and apply this to lung adenomas. By comparing the effects of the carcinogen, urethane, with those of the lung-toxic and tumor promoting agent, butylated hydroxytoluene (BHT), in mouse strains with different susceptibilities to adenoma formation, we will distinguish biochemical changes unique to neoplasia from those characteristic of a normal proliferative response. The cellular origin of the adenomas induced by urethane in adult Strain A mice is the Type 2 alveolar cell, but adenonas can also arise from Clara cells. The effect of strain, age and carcinogen on adenoma origin will be determined. A single dominant gene in BALB mice which confers resistance to adenomas will be used to differentiate toxic effects of urethane from truly neoplastic ones, and will be mapped using recombinant inbred mice. We will determine whether this gene prevents formation of hyperplastic foci or their further progression. Transplantation studies with dissected embryonic rudiments will determine if this gene acts on epithelial or mesenchymal tissue. The mechanisms by which BHT enhances adenoma formation will be studied using the sensitive analytical techniques, HPLC and GC/MS, to analyze the BHT metabolites produced in vivo and in vitro. The toxic and tumor promoting effects of BHT vary between inbred strains, species and organs. By comparing the metabolites produced in a variety of systems with the physiological effects of BHT in that system, we hope to clearly identify and distinguish between the toxic and tumor promoting derivations of this compound. Adenoma cAMP receptors have a decreased ability to bind the photoaffinity analog, 8-N3-cAMP, and have lost the ability for cAMP to stimulate autophosphorylation. Such changes will be looked for in isolated Type 2 cells at earlier stages of carcinogenesis. Antibodies will be prepared against the R-subunits of cAMP-dependent protein kinases, and used to determine changes in subunit concentrations and turnover rates. The physiological consequence of this loss of receptor sensitivity will be studied in Type 2 cells by monitoring the ability of cAMP to stimulate endogenous protein phosphorylation. Correlation between the activity of protein kinases activated by Ca++ and phospholipids in different strains and the susceptibility of these strains to adenoma induction will be thoroughly investigated, since this kinase may be involved in progression of initiated cells. A photoaffinity analog of cyclic GMP, 8-N3-(32p)cGMP, will be used to monitor the kinases activated by cyclic GMP, which change during normal development and regenerative lung cell division.

要求RCDA减轻重大的教学负荷，并允许全职 对下面描述的项目的研究以及获得经验的时间 在重组DNA技术中，并将其应用于肺腺瘤。 经过 比较致癌，氨基甲酸酯的作用 肺毒性和肿瘤促进剂，丁基化羟基甲苯（BHT） 小鼠菌株对腺瘤形成的敏感性不同，我们将 区分肿瘤特有的生化变化和 正常增殖反应的特征。 细胞来源 尿电烷在成年菌株A小鼠中诱导的腺瘤是2型 肺泡细胞，但也可能来自克拉拉细胞。 效果 将确定腺瘤起源的应变，年龄和致癌物。 一个 BALB小鼠中的主要基因，对腺瘤的抗性将是 用于区分氨基甲酸酯与真正肿瘤的毒性作用， 并将使用重组小鼠进行映射。 我们将确定 该基因是防止增生灶的形成还是进一步 进展。 通过解剖的胚胎基础的移植研究 将确定该基因是否作用于上皮组织或间质组织。 这 使用BHT增强腺瘤形成的机制将使用 敏感的分析技术HPLC和GC/MS，用于分析BHT 代谢物在体内和体外产生。 促进和肿瘤促进 BHT的影响在近交菌株，物种和器官之间有所不同。 经过 将各种系统中产生的代谢产物与 BHT在该系统中的生理影响，我们希望清楚地识别 并区分促进的毒性和肿瘤 化合物。 腺瘤营受体的结合能力降低 光性模拟，8-N3训练营，并且失去了营地的能力 刺激自磷酸化。 将以孤立的方式寻找这样的变化 在癌变的早期阶段2型细胞。 抗体将是 针对CAMP依赖性蛋白激酶的R-subunits制备，并使用 确定亚基浓度和周转率的变化。 这 这种受体敏感性丧失的生理后果将是 通过监测CAMP刺激的能力，在2型细胞中进行了研究 内源性蛋白质磷酸化。 活动之间的相关性 由Ca ++激活的蛋白激酶和不同菌株中的磷脂 这些菌株对腺瘤诱导的敏感性将是 对此进行了彻底的研究，因为该激酶可能参与进展 启动细胞。 循环GMP的光性类似物，8-N3-（32p）CGMP， 将用于监测由环状GMP激活的激酶 在正常发育和再生肺细胞分裂期间。