Microarray analysis for detecting biological characteristics in neuroblastoma

微阵列分析检测神经母细胞瘤的生物学特征

基本信息

批准号：
15209058
负责人：
HIYAMA Eiso
金额：
$ 29.12万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15209058/
关键词：
neuroblastoma microarray CGH array malignant grade caspase methylation risk assesment Taylor-made therapy hTERT 層別化

项目摘要

Microarray and array-based CGH analyses were examined in 322 human neuroblastoma cases. Array-based CGH array revealed MYCN amplification, aberrations in chromosomes 1,3,4,11,14, and 17. Custom CGH array for detection of these aberrations revealed that chromosome 11q deletion was an independently poor prognosis-associated factor. We also made custom array of 182 prognosis-related genes selected by cDNA microarray, genome-wide oligomicroarray for gene expression, and microsort technology. Expression analysis using this custom array could exhibit high accuracy (95%) to predict the neuroblastoma prognosis. We also made custom microarray for detecting DNA methylation in the promoter regions of hTERT, caspases 8 and 9. It revealed that only 2 methylated loci of caspase 8 were correlated with poor prognosis. In 9 cases with intratumoral heterogeneity, 30 tumors resected after chemotherapy, 11 multifocal tumors, and 8 recurrent tumors, 6 tumors which consequently regressed or matured, tumor c … More ells were isolated using microdissection to compare the gene expression profiling with each primary tumor. All 12 tumors whose expression profilings were extremely altered showed poor outcome, while no tumor in multifocal tumors or regressing/maturing tumors showed remarkable alteration of gene expression even if DNA ploidy patterns were different.Telomerase inhibitor or silencing of hTERT by small interfering RNA in 12 neuroblastoma cell lines resulted that 5 or 7 cell lines showed apoptosis and growth inhibition, respectively. In these cell lines, gene expression analysis revealed reduction of cell growth signals and activation of apoptosis related genes. Moreover, 3 cell lines which are induced differentiation by retinoic acid showed decrease of MYCN expression and telomerase expression and increase of apoptosis related signals and neuronal differentiation.This custom array with CGH analysis is useful to evaluate characteristics in each neuroblastoma. Moreover, it may be also a useful tool to choice the chemotherapeutic regimen and differentiation therapy. Less

在322例人神经母细胞瘤病例中检查了基于微阵列和基于阵列的CGH分析。基于阵列的CGH阵列显示MYCN扩增，染色体的畸变1,3,4,11,14和17。用于检测这些像差的自定义CGH阵列表明，11Q缺失染色体缺失是一种独立的预后较差的预后相关因子。我们还制作了由cDNA微阵列选择的182个预后相关基因的定制阵列，基因表达基因组少量阵列和微骨技术的定制阵列。使用此自定义阵列的表达分析可能存在很高的精度（95％），以预测神经母细胞瘤的预后。我们还制作了用于检测HTERT启动子区域DNA甲基化的自定义微阵列，胱天蛋白酶8和9。它表明，只有2个胱天蛋白酶8的甲基化位置与预后不良相关。在9例具有肿瘤内异质性的病例中，化学疗法后的30例肿瘤，11个多灶性肿瘤和8种复发性肿瘤，6种肿瘤，因此会退化或成熟的6个肿瘤，使用显微解剖分离出更多的ELL，以将基因表达分析与每个主要肿瘤进行比较。所有12个表达概况都极大地改变的肿瘤表现出较差的结果，而在多灶性肿瘤中没有肿瘤或回归/成熟肿瘤在基因表达上也显示出明显变化的基因表达变化，即嗜倍酶抑制剂或沉默HTERT在12个神经细胞瘤细胞中的小型rNA对HTERT的抑制作用或沉默，在5或7个细胞中都会导致5或7个细胞中的RNA产生了5或7个细胞。在这些细胞系中，基因表达分析显示细胞生长信号的降低和凋亡相关基因的激活。此外，通过视黄酸诱导分化的3种细胞系显示了MYCN表达和端粒酶表达的降低，以及凋亡相关信号和神经元分化的增加。这种具有CGH分析的自定义阵列可用于评估每个神经母细胞瘤的特征。此外，它可能也是选择化学治疗方案和分化治疗的有用工具。较少的