DNA methylation contributes to the epigenetic control of gene expression. Variations in the methylation status can result in the silencing of genes. DNA methyltransferase converts cytosine to 5-methyl cytosine in CpG islands located in the promoter regions of genes. When CpG islands are hypermethylated, the gene is repressed/silenced, and similarly when it is hypomethylated, transcription can take place and the gene is expressed. The classical methods to detect DNA methylation require labor-intensive and time-consuming steps. As a result of large-scale expression profiling studies, high-throughput techniques are needed to screen for alterations in the methylation patterns. Denaturing high performance liquid chromatography (DHPLC) is a reliable, highly sensitive technique for mutation discovery. In the present study we examined the suitability of DHPLC technology to detect alterations in methylation pattern of the promoter regions of several genes. We report reliable and reproducible results in distinguishing methylated and unmethylated promoter regions of human PCDHGB6, c-MYC, MGMT1, CDKN2A/p16, and ATM genes. These DHPLC profiles were independently confirmed with bisulfite genomic sequencing. In conclusion, DHPLC technology serves as a rapid screening tool to monitor the genomic DNA methylation and could be used to increase the throughput efficiency of methylation analysis.

中文翻译：

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用变性高效液相色谱检测基因组 DNA 甲基化。

DNA 甲基化有助于基因表达的表观遗传控制。甲基化状态的变化可导致基因沉默。DNA 甲基转移酶在位于基因启动子区域的 CpG 岛中将胞嘧啶转化为 5-甲基胞嘧啶。当 CpG 岛高度甲基化时，该基因被抑制/沉默，类似地，当它低甲基化时，可以发生转录并表达该基因。检测 DNA 甲基化的经典方法需要劳动密集型和耗时的步骤。作为大规模表达谱研究的结果，需要高通量技术来筛选甲基化模式的改变。变性高效液相色谱 (DHPLC) 是一种可靠、高度灵敏的突变发现技术。在本研究中，我们检查了 DHPLC 技术检测几个基因启动子区域甲基化模式变化的适用性。我们报告了区分人类甲基化和非甲基化启动子区域的可靠和可重复的结果。PCDHGB6、c-MYC、MGMT1、CDKN2A/p16和ATM基因。这些 DHPLC 谱通过亚硫酸氢盐基因组测序独立确认。总而言之，DHPLC 技术可作为监测基因组 DNA 甲基化的快速筛选工具，可用于提高甲基化分析的通量效率。