Mutant frequency of lacI in transgenic mice following benzo[a]pyrene treatment and partial hepatectomy

Mutat Res. 1997 Jun 9;377(1):1-11. doi: 10.1016/s0027-5107(97)00004-3.

Abstract

The Big Blue, transgenic mouse provides an in vivo mutation system that permits the study of pharmacodynamic parameters on mutant frequency (MF) following xenobiotic exposure. We have studied the effects of cellular proliferation on the frequency of mutations in the lacl transgene by evaluating the MF in the liver of male C57B1/6 Big Blue mice following treatment with benzo[a]pyrene (B[a]P) and a partial hepatectomy. Mice received either 40 mg/kg of B[a]P in corn oil or corn oil alone by i.p. injection on three consecutive days, followed by a partial hepatectomy on the fourth day. Three days later (i.e., 7 days following the initial B[a]P injection), the animals were sacrificed and the MF in the liver was compared to the MF observed in the liver of the same mouse at the time of hepatectomy. Induction of cytochrome P-450 1A (CYP1A) following B[a]P treatment was evident by Western blot analysis. The MF in untreated control animals was not significantly different at hepatectomy (4.7 +/- 0.8 x 10(-5)) and 3 days later, at sacrifice (3.0 +/- 0.4 x 10(-5)). Neither was the MF observed in the B[a]P-treated mice at the time of sacrifice (12.0 +/- 2.1 x 10(-5)) significantly different from the MF observed at the time of hepatectomy (10.6 +/- 5.3 x 10(-5)). However, B[a]P-treatment resulted in a 4.0-fold increase in MF at sacrifice which was significantly different (p < 0.05), when compared to the untreated controls. The B[a]P-treated mice at hepatectomy showed a modest 2.2-fold increase in MF which was not statistically significantly different from the untreated controls. In addition, both control and B[a]P-treated tissues gave sectored mutant plaques. The sectored plaque frequency (SPF) was significantly elevated (p < 0.05) in the B[a]P-treated mice at hepatectomy (4.2 +/- 1.0 x 10(-5)) and sacrifice (7.3 +/- 2.4 x 10(-5)) as compared to the respective frequency in the control mice at hepatectomy (1.9 +/- 0.7 x 10(-5)) and sacrifice (1.4 +/- 0.2 x 10(-5)). One explanation for this data is the persistence of the B[a]P adducts in the mouse genomic DNA that was packaged into the lambda phage, and ultimately fixed as mutations in Escherichia coli.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics*
  • Bacteriophage lambda / genetics
  • Benzo(a)pyrene / pharmacology*
  • Blotting, Western
  • Cell Division*
  • Cytochrome P-450 CYP1A1 / biosynthesis
  • Cytochrome P-450 CYP1A2 / biosynthesis
  • Enzyme Induction
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins*
  • Hepatectomy*
  • Lac Repressors
  • Liver / cytology
  • Liver / drug effects*
  • Liver / enzymology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microsomes, Liver / enzymology
  • Microsomes, Liver / metabolism
  • Mutagenicity Tests
  • Mutagens
  • Mutation*
  • RNA, Messenger / metabolism
  • Repressor Proteins / genetics*
  • Viral Plaque Assay

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Lac Repressors
  • LacI protein, E coli
  • Mutagens
  • RNA, Messenger
  • Repressor Proteins
  • Benzo(a)pyrene
  • Cytochrome P-450 CYP1A1
  • Cytochrome P-450 CYP1A2