Noninterference of cytochrome P4501A2 in the cytotoxicity of tacrine using genetically engineered V79 Chinese hamster cells for stable expression of the human or rat isoform and two human hepatocyte cell lines

Biochem Pharmacol. 1997 Feb 7;53(3):423-7. doi: 10.1016/s0006-2952(96)00713-7.

Abstract

Tacrine (THA) is the only drug currently approved for the treatment of Alzheimer's disease. A common side effect of this drug in humans is major hepatotoxicity. THA-induced toxicity may be related to a metabolic pathway implicating cytochrome P450 1A2 (CYP1A2). The purpose of this study was to clarify the role of the metabolic conversion of THA by CYP1A2 in the cytotoxicity of THA. The cytotoxicity of THA was evaluated in two human hepatocyte cell lines, HepG2 and Chang liver, and on the V79 Chinese hamster cell line, which does not express cytochrome P450 activity, and its variants, genetically engineered for expression of human or rat CYP1A2. Cells expressing human CYP1A2 metabolized THA to form its 1-OH derivative (Vmax = 9.36 +/- 0.57 pmol min(-1) mg(-1) total protein), whereas no metabolism was observed with the nonexpressing parental cells. In all cell lines, THA induced a marked decrease in cell viability and a strong inhibition of RNA and protein synthesis. However, these cytotoxic effects did not differ in parental V79 cells and variant cells expressing human or rat CYP1A2. The IC50 were tenfold higher for cell viability than for RNA and protein inhibition after 3 hr of incubation but were similar after 24 hr (P < 0.0001), indicating that this early inhibition was not a transient effect and could lead to cell death. These results strongly suggest that THA-induced cytotoxicity is not mediated by CYP1A2.

MeSH terms

  • Animals
  • Cell Line
  • Cricetinae
  • Cricetulus
  • Cytochrome P-450 CYP1A2 / genetics
  • Cytochrome P-450 CYP1A2 / physiology*
  • Genetic Engineering
  • Humans
  • Liver / cytology
  • Liver / drug effects*
  • Protein Biosynthesis
  • RNA / biosynthesis
  • Rats
  • Tacrine / toxicity*

Substances

  • Tacrine
  • RNA
  • Cytochrome P-450 CYP1A2