cyclin-d1 and Iron-Overload

Patients exhibiting hepatic iron overload frequently develop hepatocellular carcinoma. An impaired expression of hepatic genes could be involved in this phenomenon. Our aim was to identify, during iron overload, hepatic genes involved in cell cycle which are misregulated.. Mouse iron overload was obtained by carbonyl-iron supplementation or iron-dextran injection. As expected, liver iron overload was associated to both hepatomegaly and hepatocyte polyploidisation. Hepatic gene expression was investigated using macroarray hybridizations. Cyclin D1 mRNA was the only gene whose expression increased in both models. Its overexpression was confirmed by real-time quantitative PCR. Immunobloting analysis demonstrated a strong increase of Cyclin D1 protein expression in iron-overloaded hepatocytes. This overexpression was correlated with early abnormalities in their cell cycle progression judged, in vitro, on DNA synthesis and mitotic index increase.. Our data demonstrates that Cyclin D1, a protein involved in G1-phase of cell cycle, is overexpressed in the iron-overloaded liver. This iron-induced expression of Cyclin D1 may contribute to development of cell cycle abnormalities, suggesting a role of Cyclin D1 in iron-related hepatocarcinogenesis.

Topics: Animals; Blotting, Western; Cell Cycle; Cells, Cultured; Cyclin D1; Disease Models, Animal; Disease Progression; Gene Expression; Hepatocytes; Iron Overload; Liver; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Hepatomegaly is commonly observed in hepatic iron overload due to human hemochromatosis and in animal models of iron loading, but the mechanisms underlying liver enlargement in these conditions have received scant attention. In this study, male rats were treated with iron dextran or dextran alone for 6 months. Chronic iron loading resulted in a > 50-fold increase in hepatic iron concentration. Both liver weights and liver/body weight ratios were increased approximately 2-fold in the iron-loaded rats (P < 0.001 for both). Hepatocyte nuclei expressing proliferating cell nuclear antigen (PCNA), a marker of S phase, were significantly increased in the iron-loaded livers, suggesting enhanced proliferation. To assess the mechanisms by which iron promotes proliferation, the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, hepatocyte growth factor (HGF), and transforming growth factor-alpha (TGF-alpha) were assessed by reverse transcription-polymerase chain reaction (RT-PCR). Of these growth-associated factors, only TNF-alpha messenger RNA (mRNA) was significantly increased by iron loading (about 3-fold; P = 0.005). Because cyclin D1 is required for entry of hepatocytes into the cell cycle after partial hepatectomy or treatment with direct mitogens, levels of immunoreactive cyclin D1 were examined and found to be significantly increased in the iron-loaded livers. The increase in cyclin D1 protein in the iron-loaded livers was paralleled by an increase in the abundance of its transcript as measured by real-time PCR. Taken together, these results suggest that iron is a direct mitogen in the liver and raise the possibility that chronic stimulation of hepatocyte proliferation may play a role in the pathophysiology of iron overload states.

Topics: Animals; Cell Proliferation; Chronic Disease; Cyclin D1; DNA; Hepatocyte Growth Factor; Hepatocytes; Interleukin-6; Iron; Iron Overload; Liver; Male; Organ Size; Proliferating Cell Nuclear Antigen; Rats; Rats, Sprague-Dawley; RNA, Messenger; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha