Genome-wide comparison of paired fresh frozen and formalin-fixed paraffin-embedded gliomas by custom BAC and oligonucleotide array comparative genomic hybridization: facilitating analysis of archival gliomas.

Genome-wide comparison of paired fresh frozen and formalin-fixed paraffin-embedded gliomas by custom BAC and oligonucleotide array comparative genomic hybridization: facilitating analysis of archival gliomas.
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DOI:
10.1007/s00401-010-0773-z
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发表时间:
2011-04
影响因子:
12.7
通讯作者:
Louis DN
Louis DN
中科院分区:
医学1区
文献类型:
--
作者:
Mohapatra G;Engler DA;Starbuck KD;Kim JC;Bernay DC;Scangas GA;Rousseau A;Batchelor TT;Betensky RA;Louis DN

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由于基因组技术的改进和这种分析对临床肿瘤学的日益适用性,癌症的分子遗传学分析正在迅速发展。基于阵列的比较基因组杂交(aCGH)是检测DNA拷贝数改变(CNA)的有力工具,特别是在实体瘤中,并已应用于恶性胶质瘤的研究。然而,在临床环境中,神经胶质瘤通常通过小活检取样,因此福尔马林固定石蜡包埋(FFPE)块通常是可用于遗传分析的唯一组织,特别是对于罕见类型的神经胶质瘤。此外,在FFPE材料中最容易解决胶质瘤中显著瘤内异质性的生物学基础。因此,对于神经胶质瘤,使用来自FFPE组织的DNA的能力对于临床和研究应用都是必不可少的。在这项研究中,我们构建了一个定制的细菌人工染色体(BAC)阵列,并显示出良好的灵敏度和特异性检测CNA在一组配对的冷冻和FFPE胶质瘤样本。我们的研究表明,与冷冻DNA相比,FFPE中检测到的CNA之间的一致率较高。我们还开发了一种标记来自FFPE组织的DNA的方法,该方法允许与寡核苷酸阵列有效杂交。这种标记技术应用于一组双相间变性寡星形细胞瘤(AOA),以确定每个组件独特的遗传变化。总之,这些研究的结果表明,BAC和寡核苷酸aCGH是检测FFPE DNA中CNA的敏感工具,可以对罕见的、小的和/或组织学异质性胶质瘤进行全基因组分析。
Molecular genetic analysis of cancer is rapidly evolving as a result of improvement in genomic technologies and the growing applicability of such analyses to clinical oncology. Array based comparative genomic hybridization (aCGH) is a powerful tool for detecting DNA copy number alterations (CNA), particularly in solid tumors, and has been applied to the study of malignant gliomas. In the clinical setting, however, gliomas are often sampled by small biopsies and thus formalin-fixed paraffin-embedded (FFPE) blocks are often the only tissue available for genetic analysis, especially for rare types of gliomas. Moreover, the biological basis for the marked intratumoral heterogeneity in gliomas is most readily addressed in FFPE material. Therefore, for gliomas, the ability to use DNA from FFPE tissue is essential for both clinical and research applications. In this study, we have constructed a custom bacterial artificial chromosome (BAC) array and show excellent sensitivity and specificity for detecting CNAs in a panel of paired frozen and FFPE glioma samples. Our study demonstrates a high concordance rate between CNAs detected in FFPE compared to frozen DNA. We have also developed a method of labeling DNA from FFPE tissue that allows efficient hybridization to oligonucleotide arrays. This labeling technique was applied to a panel of biphasic anaplastic oligoastrocytomas (AOA) to identify genetic changes unique to each component. Together, results from these studies suggest that BAC and oligonucleotide aCGH are sensitive tools for detecting CNAs in FFPE DNA, and can enable genome-wide analysis of rare, small and/or histologically heterogeneous gliomas.
DOI: 10.1371/journal.pone.0007752
发表时间: 2009-11-13
期刊: PloS one
影响因子: 3.7
作者:
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通讯作者: Holland E
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发表时间: 2004-10-01
期刊: BIOSTATISTICS
影响因子: 2.1
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发表时间: 2008-10-23
期刊: NATURE
影响因子: 64.8
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