Phenobarbitone-induced liver response in wild type and in p53 deficient mice.

The tumour suppressor protein, p53, is involved in the regulation of apoptosis and growth arrest following DNA damage. Mutations of the p53 gene are found in 50-55% of all human cancers (Hollstein et al. Nucl. Acid Res. 22 (1994) 3551), including hepatocellular carcinomas. Phenobarbitone (PB) is a non-genotoxic hepatocarcinogen in rats and mice. With commercial availability of mice where one or both alleles of p53 have been removed we have examined the effect of PB in wild type C57BL/6J mice (p53 +/+), and p53 deficient mice (+/- and -/- p53) to determine whether p53 plays a role in the PB induced liver response. In each strain of mice, chronic administration caused liver enlargement, which was associated with centrilobular hepatocyte hypertrophy and a transient hyperplasia. In addition, an increase in centrilobular epidermal growth factor receptor and its ligand, transforming growth factor alpha and a decrease in mannose-6-phosphate receptor and its mitoinhibitory ligand, TGFbeta1 was also observed immunohistochemically. The similar response in all three strains indicates that p53 probably plays no role in the early PB induced liver effects of hypertrophy and changes in growth factor expression.

Phenobarbitone-induced ploidy changes in liver occur independently of p53.

Liver polyploidisation, characterised by accumulation of tetraploid and octaploid cells, is found with increasing age and after administration of various drugs. The significance and mechanisms controlling polyploidisation are not understood but p53 is a candidate gene to be involved. We have investigated the effect of p53 on sodium phenobarbitone (PB)-induced liver proliferation and polyploidisation. Using p53 wild type (+/+), heterozygous (+/-) and homozygous (-/-) C57BL/6J mice, we measured ploidy and proliferation (BrdU incorporation) after 21 days oral administration of PB. Administration of PB caused a striking ploidy change compared with untreated controls, with an increase in 8n cells, and no difference noted comparing the p53 genotypes. BrdU positivity also increased significantly compared with controls, with the increase in BrdU positivity occurring in 8n cells. Our results confirm that PB is a hepatic mitogen that causes liver polyploidisation with a striking increase in 8n cells within the liver. p53 status does not appear to have any effect on this PB-induced ploidy change.