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Characterization of hepcidin response to holotransferrin in novel recombinant TfR1 HepG2 cells

Research output: Contribution to journalArticlepeer-review

Kosha Mehta, Mark Busbridge, Derek Renshaw, Robert W. Evans, Sebastien Farnaud, Vinood B. Patel

Original languageEnglish
Pages (from-to)37-45
Number of pages9
JournalBlood Cells Molecules and Diseases
Volume61
Early online date30 Jun 2016
DOIs
Accepted/In press28 Jun 2016
E-pub ahead of print30 Jun 2016
PublishedOct 2016

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Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

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Abstract

Hepcidin is the key regulator of systemic iron homeostasis. The iron-sensing mechanisms and the role of intracellular iron in modulating hepatic hepcidin secretion are unclear. Therefore, we created a novel cell line, recombinant-TfR1 HepG2, expressing iron-response-element-independent TFRC mRNA to promote cellular iron-overload and examined the effect of excess holotransferrin (5g/L) on cell-surface TfR1, iron content, hepcidin secretion and mRNA expressions of TFRC, HAMP, SLC40A1, HFE and TFR2. Results showed that the recombinant cells exceeded levels of cell-surface TfR1 in wild-type cells under basal (2.8-fold; p<0.03) and holotransferrin-supplemented conditions for 24h and 48h (4.4- and 7.5-fold, respectively; p<0.01). Also, these cells showed higher intracellular iron content than wild-type cells under basal (3-fold; p<0.03) and holotransferrin-supplemented conditions (6.6-fold at 4h; p<0.01). However, hepcidin secretion was not higher than wild-type cells. Moreover, holotransferrin treatment to recombinant cells did not elevate HAMP responses compared to untreated or wild-type cells. In conclusion, increased intracellular iron content in recombinant cells did not increase hepcidin responses compared to wild-type cells, resembling hemochromatosis. Furthermore, TFR2 expression altered within 4h of treatment, while HFE expression altered later at 24h and 48h, suggesting that TFR2 may function prior to HFE in HAMP regulation.

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