STUDIES ON THE ACTIVATED FORM OF THE HUMAN PROTO-ONCOGENE, C-HA-RAS

人类原癌基因C-HA-RAS激活形式的研究

• 批准号: 3939670
• 负责人: J BRADY
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3939670
• 关键词: Adenoviridae; bladder neoplasm; clone cells; gene expression; genetic library; genetic manipulation; genetic mapping; metastasis; molecular oncology; neoplasm /cancer genetics; neoplastic transformation; oncogenes; tissue /cell culture; viral carcinogenesis; virus DNA

We have transfected various viral and cellular oncogenes into primary cultures of rat embryo cells and have obtained lines of morphologically transformed cells. Transformation with the ras oncogene alone was observed; however, a 10-fold increase in the transformation frequency was obtained when ras was cotransfected with the adenovirus E1A gene. We have examined cell lines transformed by the ras oncogene alone, and by ras plus E1A and have observed a striking difference in their metastatic potential as assayed in nude mice. Specifically, the ras alone transformants are highly metastatic, while the two gene transformants show a very low metastatic potential. Transfection of serotype 2 E1A gene, but not the serotype 12 E1A gene, into the ras alone transformants results in a substantial reduction (at least 10-fold) in the metastatic potential of these cell lines. Experiments are in progress to investigate the mechanism by which the adenovirus type 2 E1A gene reduces the metastatic potential of the ras alone transformants. In addition, we have constructed two cDNA libraries from both a high metastatic and a low metastatic cell line. These libraries will be screened with cDNA probes in order to isolate genes that are uniquely or preferentially expressed in either cell line.

我们已经将各种病毒和细胞癌基因转染到 大鼠胚胎细胞的原代培养物，并获得了 形态转化的细胞。 使用RAS进行转换 单独观察到癌基因;然而， 当ras被激活时， 与腺病毒E1 A基因共转染。 我们曾研究 ras癌基因单独转化的细胞系，以及ras + E1 A和已经观察到它们的转移性 在裸鼠中测定的潜力。 具体来说，仅ras一家 转化子是高度转移性的，而两个基因 转化体显示出非常低的转移潜能。 转染 血清2型E1 A基因，而不是血清12型E1 A基因， 单独Ras的转化体导致显著降低（在 至少10倍）。 实验正在进行中，以研究该机制， 腺病毒2型E1 A基因降低了转移性 单独的ras转化体的潜力。 另外我们有 构建了两个cDNA文库，分别来自高转移性和高转移性肿瘤。 低转移细胞系 这些文库将被筛选， cDNA探针，以分离基因， 优先在任一细胞系中表达。