title: A high-throughput and sensitive method to measure global DNA methylation: application in lung cancer.
abstract: BACKGROUND
  Genome-wide changes in DNA methylation are an epigenetic phenomenon that can lead to the development of disease. The study of global DNA methylation utilizes technology that requires both expensive equipment and highly specialized skill sets.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  By detecting these changes in global DNA methylation, CpGlobal may have a role as a barometer for the onset and development of lung cancer.
authors.affiliation: Molecular Therapeutics Division, AmberGen Incorporated, Waltham, Massachusetts, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Anisowicz
authors.forename: Hui
authors.initials: H
authors.lastname: Huang
authors.forename: Karen I
authors.initials: KI
authors.lastname: Braunschweiger
authors.forename: Ziying
authors.initials: Z
authors.lastname: Liu
authors.forename: Heidi
authors.initials: H
authors.lastname: Giese
authors.forename: Huajun
authors.initials: H
authors.lastname: Wang
authors.forename: Sergey
authors.initials: S
authors.lastname: Mamaev
authors.forename: Jerzy
authors.initials: J
authors.lastname: Olejnik
authors.forename: Pierre P
authors.initials: PP
authors.lastname: Massion
authors.forename: Richard G
authors.initials: RG
authors.lastname: Del Mastro
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001360:Avidin / Q000378:metabolism; D019921:Biotinylation; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000235:genetics*; D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D012044:Regression Analysis
chemical_list: C093483:neutravidin; D001360:Avidin