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焦点。 对于大多数人类肿瘤，没有发现这样的癌基因改变。 found. 这强烈表明，未被识别的基因被激活， 细胞遗传学和染色体杂交研究。 鳞状细胞 头颈部癌（SCCHN），多达8个新的位点似乎 特别是可能携带新的致癌基因。 尽管许多已知的癌基因不能转化NIH 3 T3细胞， 新测定法的开发受到了阻碍， 有效地抑制其他细胞类型。 腺病毒-多聚赖氨酸结合物 被发现在体外对大鼠细胞的稳定转染有效， 促进几种替代的筛选策略。 在最初的筛选中，我们分离了乳腺癌的cDNA， 腺病毒E1 a-永生化的大鼠肾细胞。 其中一个叫TAB （乳腺癌中扩增的转化基因），被发现扩增约 10-在乳腺癌MCF-7细胞中为2 - 5倍，在几种原发性乳腺癌中为2-5倍。 乳腺癌 比较全长，约400个氨基酸开放 阅读帧与数据库中的序列未能检测到相关 基因或功能基序。 TAB定位于染色体3p14.1-2 使用体细胞杂交系。 该基因被进一步定位到一个100- 使用3 p14的完整YAC重叠群，在3p14.1中的200 kb间隔处。 TAB映射 在8 Mb的区域内，该区域代表最小的 肾细胞癌基因杂合性 同样的区域， SCCHN和许多其他肿瘤。 我们建议将这种新的检测方法应用于SCCHN。 一种基于质粒的cDNA 将使用来自SCCHN细胞系的mRNA构建表达文库。 将该文库导入腺病毒E1 a永生化细胞中， 转化人cDNA将从所得转化细胞中被拯救出来， 线 这些基因将被部分测序，新的基因将被绘制出来 特定的人类染色体。 先前定位到基因座的基因 将进一步表征涉及SCCHN的特征。 这些研究的目的是 分离至少一个在SCCHN中经历体细胞改变新基因。 我们建议检测原发性SCCHN肿瘤的TAB基因扩增。 由于TAB代表与3 p14等位基因丢失相关的候选基因，我们 我建议检测SCCHN肿瘤中TAB位点的序列改变 染色体3 p缺失。