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Electrochemically Detecting DNA Methylation in the EN1 gene Promoter: Implications for understanding Ageing and DiseaseThere is a growing need for biomarkers which predict age-onset pathology. Although this is challenging, the methylome offers significant potential. Cancer is associated with the hypermethylation of many gene promoters, among which are developmental genes. Evolutionary theory suggests developmental genes arbitrate early-late life trade-offs, causing epimutations that increase disease vulnerability. Such genes could predict age related disease. The aim of this work was to optimise an electrochemical procedure for the future investigation of a broad range of ageing related pathologies. An electrochemical approach, which adopted three analytical techniques, was used to investigate DNA methylation in the EN1 gene promoter. Using synthetic single stranded DNA, one technique was able to detect DNA at concentrations as low as 10nM, with methylation status distinguishable at concentrations >25nM. A negative correlation could be observed between % methylation of heterogeneous solution and the key electrochemical parameter, Rct (r = -0.982, p < 0.01). The technique was applied to the breast cancer cell line MCF-7, where a similar correlation was observed (r = -0.965, p < 0.01). These results suggest electrochemistry can effectively measure DNA methylation at low concentrations of DNA. This has implications for the future detection of age-related disease.
The role of DNA methylation in ageing and cancerThe aim of the present review paper is to survey the literature related to DNA methylation, and its association with cancer and ageing. The review will outline the key factors, including diet, which modulate DNA methylation. Our rationale for conducting this review is that ageing and diseases, including cancer, are often accompanied by aberrant DNA methylation, a key epigenetic process, which is crucial to the regulation of gene expression. Significantly, it has been observed that with age and certain disease states, DNA methylation status can become disrupted. For instance, a broad array of cancers are associated with promoter-specific hypermethylation and concomitant gene silencing. This review highlights that hypermethylation, and gene silencing, of the EN1 gene promoter, a crucial homeobox gene, has been detected in various forms of cancer. This has led to this region being proposed as a potential biomarker for diseases such as cancer. We conclude the review by describing a recently developed novel electrochemical method that can be used to quantify the level of methylation within the EN1 promoter and emphasise the growing trend in the use of electrochemical techniques for the detection of aberrant DNA methylation.