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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Evaluation of a potential epigenetic biomarker by quantitative methyl-single nucleotide polymorphism analysis
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Evaluation of a potential epigenetic biomarker by quantitative methyl-single nucleotide polymorphism analysis

机译：通过定量甲基单核苷酸多态性分析评估潜在的表观遗传标记

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Tumorigenesis is characterized by alterations of methylation profiles including loss and gain of 5-methylcytosine. Recently, we identified a single CpG, which seemed to be consistently hypomethylated in pilocytic astrocytomas but not in other gliomas. To evaluate its applicability as a biomarker, we examined its methylation status in a large panel of gliomas (n = 97). Methylation-dependent DNA sequence variation may be considered a kind of single nucleotide polymorphism (methylSNP). MethylSNPs can be easily converted into common SNPs of the C/T type by sodium bisulfite treatment of the DNA and afterwards subjected to conventional SNP typing. We adapted SnaPshot(TM) and Pyrosequencing(TM) to determine the methylation of our test CpG in a quantitative manner. The adapted methods, called SNaPmeth and PyroMeth, respectively, gave nearly identical results, however data obtained with PyroMeth showed less scattering. Furthermore, the integrated software for allele frequency determination from Pyrosequencing could be used directly for data analysis while SnaPmeth data had to be exported and processed manually. Although data did not confirm our previous result of a preferential hypomethylation of the tested CpG in pilocytic astrocytomas, we consider quantitative methylSNP analysis by SNaPmeth or PyroMeth a favorable alternative to existing high-throughput methylation assays. It combines single CpG analysis with accurate quantitation and is amenable to high throughput. [References: 16]
机译：肿瘤发生的特征在于甲基化分布的改变，包括5-甲基胞嘧啶的丢失和增加。最近，我们鉴定出了单个CpG，在毛细胞星形细胞瘤中似乎一直被低甲基化，而在其他神经胶质瘤中却没有。为了评估其作为生物标志物的适用性，我们在一大批神经胶质瘤（n = 97）中检查了其甲基化状态。甲基化依赖的DNA序列变异可以被认为是一种单核苷酸多态性（methylSNP）。通过DNA的亚硫酸氢钠处理，可以轻松地将甲基SNP转换为C / T型常见的SNP，然后再进行常规的SNP分型。我们调整了SnaPshot（TM）和Pyrosequencing（TM），以定量的方式确定了测试CpG的甲基化程度。分别称为SNaPmeth和PyroMeth的改编方法给出了几乎相同的结果，但是用PyroMeth获得的数据显示较少的散射。此外，用于焦磷酸测序确定等位基因频率的集成软件可直接用于数据分析，而SnaPmeth数据则必须手动导出和处理。尽管数据没有证实我们先前关于在细胞性星形细胞瘤中被测CpG优先低甲基化的结果，但我们认为通过SNaPmeth或PyroMeth进行定量甲基SNP分析是现有高通量甲基化分析的有利替代方法。它结合了单CpG分析和准确的定量分析，并适合高通量。 [参考：16]

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