PILOT--ONCOGENES IN ORAL SQUAMOUS CELL CARCINOMA

试点——口腔鳞状细胞癌中的癌基因

• 批准号: 6501051
• 负责人: JEFFREY ALLEN ENGLER
• 金额: $ 26.84万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2002-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6501051
• 关键词: 3T3 cells; carcinoma; genetic library; genetic mapping; head /neck neoplasm; human genetic material tag; natural gene amplification; neoplasm /cancer genetics; neoplastic transformation; oncogenes; squamous cell carcinoma; tissue /cell culture

Identification of growth regulatory genes that are altered by mutation, gene amplification, or rearrangement is a major focus of cancer research. For the majority of human tumors, no such oncogene alteration has been found. That unidentified genes are activated is strongly suggested by cytogenetic and chromosome hybridization studies. For squamous cell carcinoma of the head and neck (SCCHN), up to eight new loci seem especially likely to harbor new oncogenes. Although many known oncogenes fail to transform NIH3T3 cells, the development of new assays has been hindered by the inability to efficiently transfect other cell types. Adenovirus-polylysine conjugate was found efficient for stable transfection of rat cells in vitro, facilitating several alternate screening strategies. In the initial screen we isolated breast cancer cDNAs that transform adenovirus E1a-immortalized rat kidney cells. One of these, named TAB (transforming gene amplified in breast cancer), was found amplified about 10-fold in breast cancer MCF-7 cells, and 2-5 fold in several primary breast cancers. Comparison of the full-length, about 400 amino acid open reading frame with sequences in the database failed to detect related genes or functional motifs. TAB was localized to chromosome 3p14.1-2 using somatic cell hybrid lines. The gene was further localized to a 100- 200kb interval in 3p14.1 using a complete YAC contig of 3p14. TAB maps within a region of 8Mb that represents the minimal region of loss of heterozygosity in renal cell carcinoma. The same region is lost from SCCHN and numerous other tumors. We propose to apply this new assay to SCCHN. A plasmid-based, cDNA expression library will be constructed using mRNA from SCCHN cell lines. The library will be introduced into adenovirus E1a-immortalized cells and transforming human cDNAs will be rescued from resulting transformed cell lines. These will be partially sequenced, and novel genes will be mapped to specific human chromosomes. Genes that map to loci previously implicated in SCCHN will be further characterized. These studies aim to isolate at least one new gene that undergoes somatic alteration in SCCHN. We propose to test primary SCCHN tumors for gene amplification of TAB. Since TAB represents a candidate gene relevant to allelic loss at 3p14, we propose to test for sequence alterations at the TAB locus in SCCHN tumors with loss of chromosome 3p.

通过突变改变的生长调节基因的鉴定 基因扩增或重排是癌症研究的主要重点。 对于大多数人类肿瘤，没有这样的癌基因改变 成立。 强烈建议，未识别的基因被激活 细胞遗传学和染色体杂交研究。 用于鳞状细胞 头部和颈部癌（SCCHN），最多八个新基因座似乎 特别有可能怀有新的癌基因。 尽管许多已知的癌基因无法转化NIH3T3细胞，但 无法开发新测定法已无法 有效转染其他细胞类型。 腺病毒 - 聚糖结合物 发现有效地在体外稳定转染大鼠细胞， 促进几种替代筛选策略。 在初始屏幕中，我们分离了转化的乳腺癌cDNA 腺病毒E1A育质大鼠肾细胞。 其中之一，命名为标签 （转化基因在乳腺癌中得到扩增），发现有关 乳腺癌MCF-7细胞中的10倍，几个主要中的2-5倍 乳腺癌。 全长的比较，约400个氨基酸开放 与数据库中序列的阅读框架未能检测到相关 基因或功能基序。 TAB被本地定位为3P14.1-2染色体 使用体细胞杂交系。 该基因被进一步定位于100- 3P14.1中的200KB间隔使用3P14的完整YAC重叠群。 标签地图 在8MB的区域内，代表最小的损失区域 肾细胞癌的杂合性。 同一地区损失了 SCCHN和许多其他肿瘤。 我们建议将此新测定法应用于SCCHN。 基于质粒的cDNA 表达式库将使用SCCHN细胞系中的mRNA构建。 该库将被引入腺病毒E1a侵蚀性细胞和 转化的人cDNA将因产生的转化细胞而被救出 线。 这些将部分测序，新的基因将被映射 特定的人类染色体。 以前映射到基因座的基因 与SCCHN有关，将进一步表征。 这些研究旨在 隔离至少一个在SCCHN中经历躯体改变的新基因。 我们建议测试原发性SCCHN肿瘤以进行TAB的基因扩增。 由于选项卡代表与3p14相关的等位基因损失相关的候选基因，我们 建议测试SCCHN肿瘤中标签基因座的序列改变 随着3p染色体的损失。