Yan Pearlly S, Wei Susan H, Huang Tim Hui-Ming
Division of Human Cancer Genetics, Department of Molecular Virology, Immunology and Molecular Genetics, The Ohio State University, Columbus, USA.
Methods Mol Biol. 2004;287:251-60. doi: 10.1385/1-59259-828-5:251.
The methylation-specific oligonucleotide (MSO) microarray is a high-throughput approach capable of detecting DNA methylation in genes across several CpG sites. Based on the bisulfite modification of DNA that converts unmethylated cytosines to uracil but leaves the 5'methylcytosine intact, the method utilizes short oligonucleotides corresponding to the methylated and unmethylated alleles as probes affixed on solid support and products amplified from bisulfite-treated DNA as targets for hybridization. MSO is suitable for examining a panel of genes across multiple clinical samples. This approach can generate a robust dataset for discovering profiles of gene methylation in cancer with aberrant DNA methylation in the neoplastic genome and widespread hypermethylation in tumor suppressor genes. MSO and other oligonucleotide-based arrays have been applied successfully for analyses of single genes and have been useful in delineating and predicting tumor subgroups using clustering methods. Here we focus on design criteria important to the interrogation of multiple CpG sites across several genes.
甲基化特异性寡核苷酸(MSO)微阵列是一种高通量方法,能够检测多个CpG位点的基因中的DNA甲基化。基于将未甲基化的胞嘧啶转化为尿嘧啶但使5'-甲基胞嘧啶保持完整的DNA亚硫酸氢盐修饰,该方法利用与甲基化和未甲基化等位基因相对应的短寡核苷酸作为固定在固体支持物上的探针,并将亚硫酸氢盐处理的DNA扩增产物作为杂交靶标。MSO适用于检测多个临床样本中的一组基因。这种方法可以生成一个强大的数据集,用于发现肿瘤中具有异常DNA甲基化的基因甲基化谱,以及肿瘤抑制基因中的广泛高甲基化。MSO和其他基于寡核苷酸的阵列已成功应用于单基因分析,并在使用聚类方法描绘和预测肿瘤亚组方面发挥了作用。在这里,我们重点关注对多个基因中的多个CpG位点进行询问时重要的设计标准。