CYP1A1 mRNA levels as a human exposure biomarker: use of quantitative polymerase chain reaction to measure CYP1A1 expression in human peripheral blood lymphocytes

Carcinogenesis. 1993 Oct;14(10):2003-6. doi: 10.1093/carcin/14.10.2003.

Abstract

Accurate human risk assessment requires sensitive methods to evaluate dose-response relationships, especially following low level exposures. We have developed a reverse transcriptase polymerase chain reaction (RT-PCR) method to quantitative cytochrome P450-1A1 (CYP1A1) mRNA levels in human blood lymphocytes. Many polycyclic aromatic hydrocarbons (PAH) such as benzo[a]pyrene, and chlorinated PAH such as polychlorinated dibenzodioxins, dibenzofurans and biphenyls induce CYP1A1 expression through activation of an endogenous protein, the Ah receptor. Using a quantitative competitive RT-PCR method that included a synthetic internal standard we determined copy numbers of CYP1A1 mRNA in resting as well as mitogen-stimulated human blood lymphocytes. In mitogen-stimulated human blood lymphocytes assay variation was approximately 10% for measurement of this low expression gene and mRNA levels correlated well with ethoxyresorufin-O-deethylase (EROD) activity. The expression of mRNA was induced 20-fold upon culturing human lymphocytes with 10 nM TCDD. In nonstimulated, uninduced lymphocytes CYP1A1 levels are extremely low (1000 copies mRNA/10(4) cells) and cannot be measured by EROD activity. Studies of CYP1A1 mRNA expression in chemically-exposed populations are in progress.

MeSH terms

  • Base Sequence
  • Biomarkers / analysis
  • Cytochrome P-450 CYP1A1
  • Cytochrome P-450 Enzyme System / analysis*
  • Environmental Exposure / analysis*
  • Humans
  • Lymphocytes / chemistry*
  • Lymphocytes / drug effects
  • Molecular Sequence Data
  • Oxidoreductases / analysis*
  • Polychlorinated Dibenzodioxins / toxicity*
  • Polymerase Chain Reaction / methods*
  • RNA, Messenger / analysis*
  • Risk Factors
  • Smoking / blood

Substances

  • Biomarkers
  • Polychlorinated Dibenzodioxins
  • RNA, Messenger
  • Cytochrome P-450 Enzyme System
  • Oxidoreductases
  • Cytochrome P-450 CYP1A1