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BMC Cancer  2008 

A high-throughput and sensitive method to measure Global DNA Methylation: Application in Lung Cancer

DOI: 10.1186/1471-2407-8-222

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Abstract:

We have designed and developed an assay, CpGlobal, which is easy-to-use, does not utilize PCR, radioactivity and expensive equipment. CpGlobal utilizes methyl-sensitive restriction enzymes, HRP Neutravidin to detect the biotinylated nucleotides incorporated in an end-fill reaction and a luminometer to measure the chemiluminescence. The assay shows high accuracy and reproducibility in measuring global DNA methylation. Furthermore, CpGlobal correlates significantly with High Performance Capillary Electrophoresis (HPCE), a gold standard technology. We have applied the technology to understand the role of global DNA methylation in the natural history of lung cancer. World-wide, it is the leading cause of death attributed to any cancer. The survival rate is 15% over 5 years due to the lack of any clinical symptoms until the disease has progressed to a stage where cure is limited.Through the use of cell lines and paired normal/tumor samples from patients with non-small cell lung cancer (NSCLC) we show that global DNA hypomethylation is highly associated with the progression of the tumor. In addition, the results provide the first indication that the normal part of the lung from a cancer patient has already experienced a loss of methylation compared to a normal individual.By detecting these changes in global DNA methylation, CpGlobal may have a role as a barometer for the onset and development of lung cancer.The functional role of DNA methylation includes maintaining the stability of chromosomes, silencing repetitive sequences, arresting the deleterious effects of integrated foreign DNA and controlling gene expression [1]. Genome-wide loss of the methyl group at 5-methyl cytosines (hypomethylation) leads to the destabilization of the DNA [2]. Global DNA hypomethylation has been observed to be one of the earliest molecular abnormalities described in human neoplasia [3,4]. This biological phenomenon could be exploited to gain an insight into the mechanism of action of DNA me

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