Techniques Used in Studies of Age-Related DNA Methylation Changes

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Epigenetic modification of CpG islands (CGIs) in promoter regions is an important regulatory mechanism of gene expression in eukaryotic cells. Hypermethylation of CGIs may silence a gene, whereas hypomethylation of previously methylated CGIs allows gene expression. The pattern of methylation is cell-type-specific and established during development of the organisms. Changes in the methylation pattern have been found in all cancer forms and in aging cells. The epigenetic-related alternations of gene expression status may significantly contribute to the initiation and maintenance of malignant growth. Cancer incidence increases dramatically with age and correlates strongly with age-related methylation changes. Many techniques have been developed to analyze the genome-wide methylation content and the methylation status of specific loci. The majority of methylation screening protocols utilizes methylation-sensitive endonuclease digestion or bisulfite treatment of the template followed by subsequent PCR amplification of a specific sequence. All methods either examine only one specific DNA sequence at a time, or provide limited genomic information on the screened sequences. The principle of our new approach is to combine methylation-sensitive enzyme digestion with the comparative genomic hybridization (CGH) technique to develop an array-based method to screen the entire genome for changes of methylation pattern. The new technique will serve as an efficient tool in understanding the nature of epigenetic changes and their significance to the aging process and cancer development.

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