Serum, plasma and paraffin-embedded tissues as sources of DNA for studying cancer susceptibility genes

Carcinogenesis. 1997 Jun;18(6):1271-5. doi: 10.1093/carcin/18.6.1271.

Abstract

The ability to isolate DNA from archived human serum, plasma and paraffin-embedded human tissues enhances opportunities to study breast, lung and other cancer risk factors. We report herein a simple and fast protocol for the extraction of genomic DNA from these sources. Using a phenol-based extraction method, the recovery for DNA is quantitative and reproducible. DNA yields in serum (250 microl) were between 162 and 1060 ng (n = 18 subjects), in plasma (250 microl) were between 165 and 375 ng (n = 5 subjects) and in embedded tissues (5-microm thick sections for ethanol fixed, and between 5- and 20-microm sections for formaldehyde fixation) were between 1 microg and 11.7 microg (n = 32 subjects). The extraction method was combined with newly designed PCR-based assays for cancer susceptibility marker genes such as CYP1A1 (exon 7), CYP2E1 (Dra1, Rsa1), GSTM1 and NAT2 [NAT2*5A (C481T), NAT2*6A (G590A), NAT2*7A (G857A)]. Genotyping results from the serum and paraffin-embedded tissues compared favorably to results from archived freshly frozen tissues, where concordance was 98% for serum, 100% for ethanol-fixed embedded tissues, and 97% for formaldehyde-fixed and paraffin-embedded tissues. This facile method will allow for the use of archived tissue samples of prospective cohort and other studies where intact DNA was not previously available.

MeSH terms

  • Arylamine N-Acetyltransferase / genetics
  • Arylamine N-Acetyltransferase / metabolism
  • Biotransformation
  • Carcinogens / pharmacokinetics
  • Cytochrome P-450 CYP1A1 / genetics
  • Cytochrome P-450 CYP1A1 / metabolism
  • Cytochrome P-450 CYP2E1 / genetics
  • Cytochrome P-450 CYP2E1 / metabolism
  • DNA / blood
  • DNA / isolation & purification*
  • DNA, Neoplasm / blood
  • DNA, Neoplasm / isolation & purification
  • Exons
  • Genetic Predisposition to Disease
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Humans
  • Inactivation, Metabolic
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Lung / chemistry
  • Lung / enzymology
  • Lung / physiology
  • Lung Neoplasms / chemistry
  • Lung Neoplasms / enzymology
  • Lung Neoplasms / genetics
  • Oncogenes*
  • Paraffin Embedding
  • Polymerase Chain Reaction / methods
  • Polymorphism, Genetic
  • Reproducibility of Results

Substances

  • Carcinogens
  • DNA, Neoplasm
  • Isoenzymes
  • DNA
  • Cytochrome P-450 CYP2E1
  • Cytochrome P-450 CYP1A1
  • Arylamine N-Acetyltransferase
  • NAT2 protein, human
  • Glutathione Transferase