Exploration of diagnostic and therapeutic targets through array-based analyses for genomic and epigenomic alterations in esophageal squamous-cell carcinoma

通过基于芯片的食管鳞状细胞癌基因组和表观基因组改变分析探索诊断和治疗靶点

基本信息

批准号：
18591457
负责人：
IMOTO Issei
金额：
$ 2.53万
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：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

By constructing the array-based system for genomic, epigenomic, and mRNA and protein expression changes, we tried to identify diagnostic and/or therapeutic target genes necessary and sufficient to develop "personalized medicine" for esophageal squamous-cell carcinoma (ESCC).(1) Investigation of genomic copy-number aberrations using CGH-array in ESCC In cell lines as well as primary tumor samples of ESCC, we performed array-CGH analyses using bacterial artificial chromosome (BAC)-arrays and oligo-arrays to explore novel copy-number alterations in cancer genome. Among series of genes locating within the identified amplified and homozygously deleted regions, we determined several target genes, which are associated with the pathogenesis of ESCC through these alterations. In addition to the ESCC, we also analyzed several types of cancers, such as ovarian cancer, hepatocellular carcinoma, and oral cancers, and identified several targets, which may be useful for diagnosis/prediction of progno … More sis and molecular targeting therapy.(2) Evaluation of clinicopathological and biological significance of candidate targets in cancer cells Possible cancer-associated genes identified from amplified and homozygously deleted regions, we performed expression analyses in mRNA and protein levels using quantitative methods, such as array-based methods and real-time PCR system. In candidate tumor-suppressor genes (TSGs), we also analyzed the involvement of epigenetic alterations, such as alterations in DNA methylation and histone codes, in cancer cells. To evaluate biological significance of target genes as oncogenes or TSGs, we transfected expression plasmids or siRNAs for those genes, and analyzed cell growth, cell cycle, apoptotic cell death, and/or invasive phenotype.(3) Identification of TSGs through direct exploration of epigenetic alterations in cancer In order to identify epigenetically silenced genes directly, we applied BAMCA (BAC-array based methylated CpG island amplification) method to cancer DNA, and explored abnormally methylated region. From identified possibly hypermethylated regions in ESCC cell lines, we identified several candidate TSGs and determined their clinicopathological and biological significance in ESCC. Less

By constructing the array-based system for genomic, epigenomic, and mRNA and protein expression changes, we tried to identify diagnostic and/or therapeutic target genes necessary and sufficient to develop "personalized medicine" for esophageal squamous-cell carcinoma (ESCC).(1) Investigation of genomic copy-number aberrations using CGH-array in ESCC In cell lines as well as primary tumor samples of ESCC, we performed Array-CGH使用细菌人工染色体（BAC）和寡聚阵列进行分析，以探索癌症基因组中新型的拷贝数改变。在位于已鉴定的扩增和纯合区域中的一系列基因中，我们确定了几个靶基因，这些基因与ESCC的发病机理通过这些改变有关。除ESCC外，我们还分析了几种类型的癌症，例如卵巢癌，肝细胞癌和口服癌症，并确定了几个靶标，这些靶标可能对Progno…的诊断/预测可能有用。纯合删除的区域，我们使用基于阵列的方法和实时PCR系统等定量方法在mRNA和蛋白水平上进行了表达分析。在候选肿瘤 - 抑制基因（TSG）中，我们还分析了表观遗传学改变的参与，例如癌细胞中DNA甲基化和组蛋白代码的改变。 To evaluate biological significance of target genes as oncogenes or TSGs, we translated expression plasmids or siRNAs for those genes, and analyzed cell growth, cell cycle, apoptotic cell death, and/or invasive phenotype.(3) Identification of TSGs through direct exploration of epigenetic alterations in cancer In order to identify epigenetically silenced genes directly, we applied BAMCA (BAC-array based methylated CpG岛扩增）癌症DNA的方法，并探索了绝对甲基化区域。根据ESCC细胞系中可能的高甲基化区域，我们确定了几个候选TSG，并确定了它们在ESCC中的临床病理学和生物学意义。较少的