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

• 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

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)

我们构建了基于阵列的系统，用于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的方式筛选推定的ESCC相关基因。1）使用阵列比较基因组杂交分析基因组拷贝数畸变我们分析了来自ESCC细胞系的基因组DNA，以及使用内部阵列-CGH通过显微切割特异性分离的原发性肿瘤细胞。阵列计算全息图（Array-CGH）我们试图从那些表现出隐性但显著的基因组拷贝数变化的区域，如高水平扩增和纯合缺失，来鉴定这些畸变的靶基因。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志物，如淋巴结转移和预后。将同样的mRNA和蛋白质表达分析方法应用于其他癌症，导致许多癌症相关基因的鉴定，包括与肿瘤相关的基因。2）使用基于BAC阵列的甲基化CpG岛扩增（BAMCA）鉴定因DNA甲基化而沉默的肿瘤抑制基因我们将MCA方法与阵列CGH（BAMCA）相结合，并筛选出癌症中异常的甲基化序列。使用BAMCA与以下数据库搜索和表达分析，有和没有药理学修饰，我们成功地确定了ESC和其他肿瘤中的甲基化沉默的几个基因。其中，有一个基因在胚胎干细胞中经常被CpG岛甲基化所沉默，并对胚胎干细胞表现出生长抑制作用，提示该基因是胚胎干细胞的候选抑癌基因（未发表数据）

项目成果

Involvement of cyclin D3 in liver metastasis of colorectal cancer, revealed by genome-wide copy-number analysis

• DOI: 10.1038/labinvest.3700312
• 发表时间: 2005-09
• 期刊: Laboratory Investigation
• 影响因子: 5
• 作者: H. Tanami;H. Tsuda;S. Okabe;T. Iwai;K. Sugihara;I. Imoto;J. Inazawa

Frequent silencing of the candidate tumor suppressor PCDH20 by epigenetic mechanism in non-small-cell lung cancers

• DOI: 10.1158/0008-5472.can-05-4437
• 发表时间: 2006-05-01
• 期刊: CANCER RESEARCH
• 影响因子: 11.2
• 作者: Imoto, Issei;Izumi, Hiroyuki;Inazawa, Johji
• 通讯作者: Inazawa, Johji

Overexpression of PDZK1 within the 1q12-q22 amplicon is likely to be associated with drug resistance phenotype in multiple myeloma

• DOI: 10.1016/s0002-9440(10)63276-2
• 发表时间: 2004-07-01
• 期刊: AMERICAN JOURNAL OF PATHOLOGY
• 影响因子: 6
• 作者: Inoue, J;Otsuki, T;Inazawa, J
• 通讯作者: Inazawa, J

Frequent silencing of low density lipoprotein receptor-related protein 1B (LRP1B) expression by genetic and epigenetic mechanisms in esophageal squamous cell carcinoma

• DOI: 10.1158/0008-5472.can-04-0172
• 发表时间: 2004-06-01
• 期刊: CANCER RESEARCH
• 影响因子: 11.2
• 作者: Sonoda, I;Imoto, I;Inazawa, J
• 通讯作者: Inazawa, J

Overexpressed Skp2 within 5p amplification detected by array‐based comparative genomic hybridization is associated with poor prognosis of glioblastomas

• DOI: 10.1111/j.1349-7006.2005.00099.x
• 发表时间: 2005-10
• 期刊: Cancer Science
• 影响因子: 5.7
• 作者: K. Saigusa;N. Hashimoto;H. Tsuda;S. Yokoi;M. Maruno;T. Yoshimine;M. Aoyagi;Kikuo Ohno;I. Imoto;J. Inazawa