Unidirectional upregulation of the synthesis of the major iron proteins, transferrin-receptor and ferritin, in HepG2 cells by the acute-phase protein α1-antitrypsin

Background/Aims: We have previously shown that the hepatic acute-phase protein α1-antitrypsin (α1-AT) is an important mediator of changes in iron metabolism in the course of anaemia of chronic disease. α1-AT profoundly reduces growth of erythroid cells by interfering with transferrin-mediated iron uptake. In the present work we investigate the effects of α1-AT on hepatic iron metabolism, as the liver plays a central role in body iron metabolism and in metabolic changes during acute-phase response. Methods: The human hepatoma cell line Hep G2 was cultured in RPMI 1640+10% FCS. The effect of α1-AT on transferrin-receptor binding was investigated in equilibrium binding assays with 125I-transferrin. Expression of transferrin receptor was determined by saturation experiments and intracellular ferritin was measured in cell lysates after incubating cells either alone or with α1-AT. To determine iron regulatory protein binding activity to iron responsive elements we used gel retardation assays and Northern blot analysis was carried out to investigate transferrin receptor and ferritin mRNA expression. Results: α1-AT completely prevented transferrin from binding to its receptor and internalization of the transferrin-transferrin receptor complex on HepG2. In addition, α1-AT caused a distinct increase in iron regulatory protein binding activity to iron responsive elements, which is characteristic of iron deprivation. Normally, the synthesis of transferrin receptor and ferritin is regulated bidirectionally, but α1-AT promoted a unidirectional regulation. α1-AT increased the synthesis of both transferrin receptor and ferritin and, moreover, increased cellular amounts of transferrin receptor mRNA and ferritin H-chain mRNA. Conclusions: The effect of α1-AT on transferrin receptor synthesis appears to be mediated via activation of iron responsive element binding affinity of iron regulatory protein leading to an increased stability of transferrin receptor mRNA. Changes in ferritin, however, may be related to a transcriptionally mediated, iron-independent pathway which overrides the influence of activated iron regulatory protein. These specific changes in iron metabolism are the very ones seen in the course of anaemia of chronic disease. Our results emphasize the central role of α1-AT as a mediator of altered iron metabolism, characteristic of anaemia of chronic disease, not only with respect to erythroid cells but also with respect to liver cells.