Identification of molecular target for diagnosis and therapy of esophageal squamous-cell carcinoma based on microarray technology.

基于微阵列技术识别食管鳞癌诊治分子靶点

基本信息

批准号：
16591300
负责人：
IMOTO Issei
金额：
$ 2.3万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591300/
关键词：
esophageal cancer array-CGH genome epigenome DNA methylation tissue microarray expression analysis molecular target CGHアレイ

项目摘要

We have constructed array-based system to analyze genomic DNA copy-number, mRNA expression, and protein expression quantitatively and qualitatively. In order to identify a set of genes useful for the molecular-targeted diagnosis and therapy of esophageal squamous-cell carcinoma (ESCC), we applied our system to screen putative ESCC-associate genes in a genome-wide manner.1) Analysis of genomic copy-number aberration using array-comparative genomic hybridization (array-CGH) and identification of target geneWe analyzed genomic DNA from ESCC cell lines as well as primary tumor cells specifically isolated by microdissection using in-house array-based CGH (array-CGH). From the regions showing cryptic but remarkable genomic copy-number changes, such as high-level amplification and homozygous deletion, we tried to identify target genes for those aberrations. In addition, we explored molecular markers associated with specific phenotypes, such as lymph node metastasis and prognosis, by comparing pattern of genomic copy-number changes and clinicopathological parameters. The same approach with mRNA and protein expression analyses was applied to other cancers, resulting in the identification of many cancer-related genes, including genes correlated with prognosis.2) Identification of tumor-suppressor genes silenced by DNA methylation using BAC-array-based methylated CpG-island amplification (BAMCA)We combined MCA method to amplify methylated sequences with array-CGH (BAMCA), and screened aberrantly methylated sequences in cancers. Using BAMCA with following database search and expression analyses with and without pharmacological modifications, we successfully identified several genes silenced by methylation in ESC and other tumors. Among them, one gene was frequently silenced by CpG island methylation in ESC and showed growth suppressive effect on ESC cells, suggesting that this gene is a candidate tumor-suppressor for ESC (unpublished data)

我们构建了基于芯片的系统来定量和定性分析基因组 DNA 拷贝数、mRNA 表达和蛋白质表达。为了鉴定一组可用于食管鳞状细胞癌（ESCC）分子靶向诊断和治疗的基因，我们应用我们的系统以全基因组方式筛选假定的食管鳞状细胞癌相关基因。1）基因组分析使用阵列比较基因组杂交 (array-CGH) 进行拷贝数畸变并鉴定靶基因我们使用内部基于阵列的 CGH 分析了来自 ESCC 细胞系以及通过显微切割专门分离的原发性肿瘤细胞的基因组 DNA （数组-CGH）。从显示神秘但显着的基因组拷贝数变化的区域，例如高水平扩增和纯合删除，我们试图识别这些畸变的目标基因。此外，我们通过比较基因组拷贝数变化模式和临床病理参数，探索与特定表型相关的分子标记，例如淋巴结转移和预后。同样的 mRNA 和蛋白质表达分析方法也适用于其他癌症，从而鉴定出许多癌症相关基因，包括与预后相关的基因。2) 使用基于 BAC 阵列的方法鉴定因 DNA 甲基化而沉默的肿瘤抑制基因甲基化CpG岛扩增（BAMCA）我们将MCA方法与array-CGH（BAMCA）相结合来扩增甲基化序列，并筛选癌症中的异常甲基化序列。使用 BAMCA 进行以下数据库搜索和表达分析（有或没有药理学修饰），我们成功地鉴定了 ESC 和其他肿瘤中因甲基化而沉默的几个基因。其中，一个基因在ESC中经常被CpG岛甲基化沉默，并对ESC细胞表现出生长抑制作用，表明该基因是ESC的候选肿瘤抑制基因（未发表数据）