EFFECT OF MYC-GENE EXPRESSION ON GROWTH IN LUNG CANCER

MYC 基因表达对肺癌生长的影响

• 批准号: 3180745
• 负责人: GEORGE D. SORENSON
• 金额: $ 12.42万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 1988-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3180745
• 关键词: adrenocorticotropic hormone; aromatic L aminoacid decarboxylase; calcitonin; clone cells; gene expression; genetic manipulation; hormone biosynthesis; human tissue; molecular cloning; molecular oncology; natural gene amplification; neoplasm /cancer genetics; neoplastic growth; oncogenes; small cell lung cancer; adrenocorticotropic hormone; aromatic L aminoacid decarboxylase; calcitonin; clone cells; gene expression; genetic manipulation; hormone biosynthesis; human tissue; molecular cloning; molecular oncology; natural gene amplification; neoplasm /cancer genetics; neoplastic growth; oncogenes; small cell lung cancer

The object of this project is to evaluate the role of myc-gene expression in influencing biological and biochemical characteristics of small carcinoma of the lung (SCCL). SCCL can be histopathologically categorized into at least two forms. The predominant form(s) have long been recognized as the small (oat) cell and intermediate subtypes. Another less common form is the mixed small cell/large cell morphologic type. Recently continuous cultures of SCCL have also been divided into two major forms, i.e., 1) the classic form and 2) the variant form. These two types have different biochemical and biological characteristics and have been correlated with the small cell/intermediate cell and small cell/large cell histopathologic forms in patients respectively. Of particular interest has been the observation that practically all the variant cell lines have significant amplification of the c-myc or the related n-myc genes while this rarely occurs in the classic cell lines. The specific aims of this investigation will be to: 1) cotransfect SCCL cells which are not expressing the c-myc gene with cloned v-myc or c-myc sequences and vector-Ecogpt DNA using the calcium phosphate transfection method; 2) select cells containing the transfected genes with the mycophenolic acid selection method; 3) clone the transfected cells and determine the relative expression of the myc gene in these transfected clones by Southern and Northern blotting of DNA and RNA respectively and hybridization with a c-myc probe; and 4) compare the populations of cells containing the transfected activated myc-gene for: (a) proliferative capacity; (b) colony forming capability; (c) tumorigenicity in nude mice; and (d) hormone as well as L-DOPA decarboxylase production with the original cell line which did not express the myc-gene. These investigations will lead to a much better understanding of the role of c-myc expression in SCCL and whether or not it is significantly related to important biological and functional characteristics of this tumor. (X)

该项目的目的是评估Myc-Gene表达的作用 在影响小的生物学和生化特征方面 肺癌（SCCL）。 SCCL可以在组织病理学上分类 至少两种形式。 长期以来一直认识到主要形式 作为小（燕麦）细胞和中间亚型。 另一个不太常见 形式是混合的小细胞/大细胞形态类型。 最近 SCCL的连续培养也已分为两种主要形式： 即1）经典形式和2）变体形式。 这两种类型有 不同的生化和生物学特征 与小细胞/中间细胞和小细胞/大细胞相关 患者的组织病理学形式分别。 特别感兴趣的 一直以来，几乎所有变体细胞系都有 C-MYC或相关N-MYC基因的显着扩增，而 这很少发生在经典细胞系中。 这项调查的具体目的是：1）共转化SCCL 未用克隆的V-MYC或C-MYC表达C-MYC基因的细胞 使用磷酸钙转染序列和载体-Ecogpt DNA 方法; 2）选择包含带有转染基因的细胞 霉酚酸选择方法； 3）克隆转染的细胞和 确定在这些转染中MYC基因的相对表达 DNA和RNA的南部和北部印迹的克隆以及 与C-MYC探针杂交； 4）比较细胞的种群 包含转染的激活的myc-gene：（a）增殖 容量; （b）菌落形成能力； （c）裸鼠的致瘤性； （d）激素以及L-DOPA脱羧酶的产生 原始细胞系未表达Myc-Gene。 这些调查将导致对角色的更好理解 SCCL中C-MYC表达的表达以及它是否显着相关 对于该肿瘤的重要生物学和功能特征。 （x）