Expression patterns of cell cycle proteins in the livers of rats treated with hepatocarcinogens for 28 days

• Published in: Archives of toxicology 2013-06, Vol.87 (6), p.1141-1153
• Main Authors: Yafune, Atsunori, Taniai, Eriko, Morita, Reiko, Nakane, Fumiyuki, Suzuki, Kazuhiko, Mitsumori, Kunitoshi, Shibutani, Makoto
• Format: Article
• Language: English
• Subjects: Acetaminophen; Animals; Apoptosis - drug effects; Biomedical and Life Sciences; Biomedicine; Carcinogens; Carcinogens - toxicity; Cell cycle; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell Proliferation - drug effects; Environmental Health; Gene Expression Profiling - methods; Gene Expression Regulation, Neoplastic - drug effects; Genotoxicity and Carcinogenicity; Immunohistochemistry; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Kruppel-Like Factor 6; Kruppel-Like Transcription Factors - genetics; Kruppel-Like Transcription Factors - metabolism; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver Neoplasms - chemically induced; Liver Neoplasms - genetics; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; M Phase Cell Cycle Checkpoints - drug effects; Male; Occupational Medicine/Industrial Medicine; Oligonucleotide Array Sequence Analysis; Organ Size - drug effects; Pharmacology/Toxicology; Proteins; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - metabolism; Proto-Oncogene Proteins c-mdm2 - genetics; Proto-Oncogene Proteins c-mdm2 - metabolism; Rats; Rats, Inbred F344; Real-Time Polymerase Chain Reaction; Rodents; Time Factors; Toxicology; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism
• ISSN: 0340-5761; 1432-0738
• DOI: 10.1007/s00204-013-1011-y

Some hepatocarcinogens induce cytomegaly, which reflects aberrant cell cycling and increased ploidy, from the early stages of administration to animals. To clarify the regulatory molecular mechanisms behind cell cycle aberrations related to the early stages of hepatocarcinogenesis, we performed gene expression analysis using microarrays and real-time reverse transcription polymerase chain reaction followed by immunohistochemical analysis in the livers of rats treated with the cytomegaly inducing hepatocarcinogens thioacetamide (TAA), fenbendazole, and methyleugenol, the cytomegaly non-inducing hepatocarcinogen piperonyl butoxide (PBO), or the non-carcinogenic hepatotoxicants acetaminophen and α -naphthyl isothiocyanate, for 28 days. Gene expression profiling showed that cell cycle-related genes, especially those of G 2 /M phase, were mostly upregulated after TAA treatment. Immunohistochemical analysis was performed on cell cycle proteins that were upregulated by TAA treatment and on related proteins. All hepatocarcinogens, irrespective of their cytomegaly inducing potential, increased liver cells immunoreactive for p21 Cip1 , which acts on cells arrested in G 1 phase, and for Aurora B or Incenp, which is suggestive of an increase in a cell population with chromosomal instability caused by overexpression. PBO did not induce cell proliferation after 28-day treatment. Hepatocarcinogens that induced cell proliferation after 28-day treatment also caused an increase in p53 + cells in parallel with increased apoptotic cells, as well as increased population of cells expressing M phase-related proteins nuclear Cdc2, phospho-Histone H3, and HP1 α . These results suggest that hepatocarcinogens may increase cellular populations arrested in G 1 phase or showing chromosomal instability after 28-day treatment. Hepatocarcinogens that induce cell cycle facilitation may cause M phase arrest accompanied by apoptosis.