Lips Esther H, Dierssen Jan Willem F, van Eijk Ronald, Oosting Jan, Eilers Paul H C, Tollenaar Rob A E M, de Graaf Eelco J, van't Slot Ruben, Wijmenga Cisca, Morreau Hans, van Wezel Tom
Department of Pathology, Leiden University Medical Center, the Netherlands.
Cancer Res. 2005 Nov 15;65(22):10188-91. doi: 10.1158/0008-5472.CAN-05-2486.
Most human cancers show genetic instabilities leading to allelic imbalances, including loss of heterozygosity (LOH). Single nucleotide polymorphism (SNP) arrays can be used to detect LOH. Currently, these arrays require intact genomic DNA as obtained from frozen tissue; however, for most cancer cases, only low-quality DNA from formalin-fixed, paraffin-embedded (FFPE) tissue is available. In this study, we tested Illumina BeadArrays to genotype FFPE tissue and detect LOH/allelic imbalances in matched colorectal tumor and normal tissue. Genotypes were compared between leukocyte and FFPE normal tissue as well as between frozen and FFPE tumor tissue. Identical genotypes and LOH profiles were obtained from normal and tumor isolates. LOH was mainly observed on chromosomes 4, 5q, 12q, 14q, 15q, 17p, 18, and 20p, which are commonly detected regions in colorectal cancer. LOH profiles of the BeadArrays were compared with profiles obtained by Affymetrix GeneChip 10K arrays, showing identical LOH patterns. These data show that genome-wide genotyping of FFPE tissue with the BeadArray gives reliable results and is a powerful technique for LOH analysis.
大多数人类癌症表现出导致等位基因失衡的基因不稳定,包括杂合性缺失(LOH)。单核苷酸多态性(SNP)阵列可用于检测LOH。目前,这些阵列需要从冷冻组织中获得的完整基因组DNA;然而,对于大多数癌症病例,只有来自福尔马林固定、石蜡包埋(FFPE)组织的低质量DNA可用。在本研究中,我们测试了Illumina BeadArrays对FFPE组织进行基因分型,并检测匹配的结直肠肿瘤组织和正常组织中的LOH/等位基因失衡。比较了白细胞与FFPE正常组织之间以及冷冻与FFPE肿瘤组织之间的基因型。从正常和肿瘤分离物中获得了相同的基因型和LOH谱。LOH主要出现在4号、5q、12q、14q、15q、17p、18号和20p染色体上,这些是结直肠癌中常见的检测区域。将BeadArrays的LOH谱与Affymetrix GeneChip 10K阵列获得的谱进行比较,显示出相同的LOH模式。这些数据表明,使用BeadArray对FFPE组织进行全基因组基因分型可获得可靠结果,并且是一种用于LOH分析的强大技术。