PILOT--ONCOGENES IN ORAL SQUAMOUS CELL CARCINOMA

试点——口腔鳞状细胞癌中的癌基因

• 批准号: 6472276
• 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: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2002-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6472276
• 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.

鉴定因突变而改变的生长调节基因， 基因扩增或重排是癌症研究的一个主要焦点。 对于大多数人类肿瘤来说，没有这样的癌基因改变 找到了。未知基因被激活的强烈建议是 细胞遗传学和染色体杂交研究。对于鳞状细胞 头颈部癌(SCCHN)，多达8个新的基因位点 尤其可能存在新的致癌基因。 尽管许多已知的癌基因无法转化NIH3T3细胞，但 新的分析方法的发展一直受到阻碍，因为无法 高效地转化其他类型的细胞。腺病毒-多聚赖氨酸结合物 在体外对大鼠细胞的稳定转染是有效的， 促进了几种可供选择的筛选策略。 在最初的筛查中，我们分离了乳腺癌的cDNA 腺病毒E1a-永生化大鼠肾细胞。其中之一，名为TAB (转化基因在乳腺癌中扩增)，被发现扩增了约 在乳腺癌MCF-7细胞中是10倍，在几种原代细胞中是2-5倍 乳腺癌。比较全长，约400个氨基酸开放 与数据库中的序列相关联的读框检测失败 基因或功能模体。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100- 3p14.1中的200kb间隔，使用3p14的完整YAC重叠群。选项卡式地图 在代表最小损耗区域的8Mb区域内 肾细胞癌的杂合性。同样的地区也从 SCCHN和许多其他肿瘤。 我们建议将这一新的检测方法应用于SCCHN。一种以质粒为基础的基因 表达文库将使用SCCHN细胞系的mRNA构建。 该文库将被引入腺病毒E1a永生化细胞中，并 转化的人类DNA将从转化的细胞中解救出来 台词。这些基因将被部分测序，新的基因将被绘制出来 人类特定的染色体上。以前映射到基因座的基因 与SCCHN有牵连的人将被进一步定性。这些研究旨在 分离至少一个在SCCHN中经历体细胞变化的新基因。 我们建议对原发SCCHN肿瘤进行TAB基因扩增检测。 由于TAB代表与3p14等位基因丢失相关的候选基因，我们 建议测试SCCHN肿瘤中TAB基因的序列变化 伴随3p染色体的丢失。