Microarray analysis for detecting biological characteristics in neuroblastoma

对322例神经母细胞瘤患者进行了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1、3、4、11、14和17的畸变。定制CGH阵列检测这些畸变显示，染色体11 q缺失是一个独立的不良预后相关因素。我们还定制了182个与乳腺癌相关的基因，这些基因是通过cDNA微阵列、全基因组基因表达寡核苷酸微阵列和微分选技术筛选出来的。使用这种定制阵列的表达分析可以表现出高准确性（95%）来预测神经母细胞瘤预后。我们还制作了用于检测hTERT、半胱天冬酶8和9的启动子区域中的DNA甲基化的定制微阵列。结果表明，caspase 8基因仅有2个甲基化位点与预后不良相关。在9例肿瘤内异质性的病例中，30例肿瘤在化疗后切除，11例为多灶性肿瘤，8例为复发性肿瘤，6例肿瘤因此而消退或成熟，11例为复发性肿瘤。 关于我们 使用显微切割分离细胞以比较每个原发性肿瘤的基因表达谱。所有12个肿瘤的表达谱发生了极大的变化，显示了不良的结果，而肿瘤的多灶性肿瘤或退化/成熟的肿瘤显示，即使DNA倍体模式不同的基因表达的显着变化。端粒酶抑制剂或沉默hTERT在12个神经母细胞瘤细胞株，分别有5个或7个细胞株出现凋亡和生长抑制。在这些细胞系中，基因表达分析显示细胞生长信号减少和凋亡相关基因激活。此外，经维甲酸诱导分化的3株神经母细胞瘤细胞，MYCN和端粒酶的表达均降低，凋亡相关信号和神经元分化增强。此外，它也可能是一个有用的工具，选择化疗方案和分化治疗。少

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