Cyp1a1 mrna levels as a human exposure biomarker: Use of quantitative polymerase chain reaction to measure cyp1a1 expression in human peripheral blood lymphocytes

John Patrick Vanden Heuvel, George C. Clark, Claudia L. Thompson, Zadock Mccoy, Chris R. Miller, George W. Lucier, Douglas A. Bell

Research output: Contribution to journalArticle

126 Scopus citations

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 quantitate 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 CYP1AI 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 lympocytes assay variation was ∼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.

Original languageEnglish (US)
Pages (from-to)2003-2006
Number of pages4
JournalCarcinogenesis
Volume14
Issue number10
DOIs
Publication statusPublished - Oct 1 1993

    Fingerprint

All Science Journal Classification (ASJC) codes

  • Cancer Research

Cite this