Correlation of LINE-1 methylation levels in patient-matched buffy coat, serum, buccal cell, and bladder tumor tissue DNA samples

Cancer Epidemiol Biomarkers Prev. 2012 Jul;21(7):1143-8. doi: 10.1158/1055-9965.EPI-11-1030. Epub 2012 Apr 26.

Abstract

Background: Evidence suggests that global methylation levels in blood cell DNA may be a biomarker for cancer risk. To date, most studies have used genomic DNA isolated from blood or urine as a surrogate marker of global DNA methylation levels in bladder tumor tissue.

Methods: A subset of 50 bladder cancer cases was selected from the New England Bladder Cancer Case-Control Study. Genomic DNA was isolated from buffy coat, buccal cells, serum, and formalin-fixed, paraffin-embedded tissue for each participant. DNA methylation at four CpG sites within the long interspersed nucleotide element (LINE-1) repetitive element was quantified using pyrosequencing and expressed as a mean methylation level across sites.

Results: Overall, the mean percent (%) LINE-1 5-methylcytosine (%5MeC) level was highest in serum (80.47% ± 1.44%) and lowest in bladder tumor DNA (61.36% ± 12.74%) and levels varied significantly across tissue types (P = 0.001). An inverse association between LINE-1 mean %5MeC and tumor stage (P = 0.001) and grade (P = 0.002) was observed. A moderate correlation between patient-matched serum and buffy coat DNA LINE-1 %5MeC levels was found (r = 0.32, P = 0.03) but levels were uncorrelated among other matched genomic DNA samples.

Conclusions: The mean promoter LINE-1 %5MeC measurements were correlated between buffy coat and serum DNA samples. No correlation was observed between genomic DNA sources and tumor tissues; however a significant inverse association between tumor percent LINE-1 methylation and tumor stage/grade was found.

Impact: LINE-1 methylation measured in case blood DNA did not reflect that observed in bladder tumor tissue but may represent other factors associated with carcinogenesis.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Aged
  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / genetics*
  • Blood Cells / metabolism*
  • Blood Cells / pathology
  • Case-Control Studies
  • DNA / genetics
  • DNA Methylation*
  • Female
  • Humans
  • Long Interspersed Nucleotide Elements / genetics*
  • Male
  • Middle Aged
  • Mouth Mucosa / metabolism*
  • Mouth Mucosa / pathology
  • Neoplasm Grading
  • Neoplasm Staging
  • Polymerase Chain Reaction
  • Prognosis
  • Survival Rate
  • Urinary Bladder / metabolism*
  • Urinary Bladder / pathology
  • Urinary Bladder Neoplasms / diagnosis
  • Urinary Bladder Neoplasms / genetics*
  • Urinary Bladder Neoplasms / mortality

Substances

  • Biomarkers, Tumor
  • DNA