Gao L, Cheng L, Zhou J N, Zhu B L, Lu Z H
Key Laboratory for Molecular and Biomolecular Electronics of Ministry of Education, Southeast University, Nanjing 210096, China.
Colloids Surf B Biointerfaces. 2005 Feb 25;40(3-4):127-31. doi: 10.1016/j.colsurfb.2004.10.024.
We described a DNA microarray-based method combined with bisulphite treatment of DNA and regular PCR to examine hyper-methylation in promoter 1A of APC gene. A set of oligonucleotide probes were designed and immobilized on the aldehyde-coated glass slides for detecting the methylation pattern of 15 selected CpG sites in the region. The methylation status of 30 colorectal tumor samples have been examined by both of methylation-specific PCR (MS-PCR) and the present microarray method. The methylation pattern of the 15 CpG sites for the samples have been obtained with the microarray. A total of 19 samples out of 30 were methylated by microarray, in which five samples cannot be detected by MS-PCR due to the methylated CpG patterns not accordant to the MS-PCR primers. The detecting ratio for methylation of APC gene of colorectal tumor samples increased from 46.7% with MS-PCR to 63.3% with the microarray, which successfully demonstrated that DNA microarray-based method not only can obtained the methylation patterns for the related genes, but also decrease the false-negative results of methylation status by the conventional MS-PCR for the investigated genes.
我们描述了一种基于DNA微阵列的方法,该方法结合了DNA的亚硫酸氢盐处理和常规PCR,用于检测APC基因启动子1A中的高甲基化。设计了一组寡核苷酸探针,并将其固定在醛包被的载玻片上,以检测该区域中15个选定CpG位点的甲基化模式。通过甲基化特异性PCR(MS-PCR)和本微阵列方法对30个结直肠肿瘤样本的甲基化状态进行了检测。用微阵列获得了样本中15个CpG位点的甲基化模式。30个样本中共有19个样本通过微阵列检测为甲基化,其中5个样本由于甲基化的CpG模式与MS-PCR引物不一致而无法通过MS-PCR检测到。结直肠肿瘤样本中APC基因甲基化的检测率从MS-PCR的46.7%提高到微阵列的63.3%,这成功地证明了基于DNA微阵列的方法不仅可以获得相关基因的甲基化模式,而且可以减少传统MS-PCR对所研究基因甲基化状态的假阴性结果。