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

Mehta, K., Busbridge, Mark, Renshaw, D., Evans, R.W., Farnaud, S. and Patel, V. 2016. Characterization of hepcidin response to holotransferrin in novel recombinant TfR1 HepG2 cells. Blood Cells, Molecules, and Diseases. 61, pp. 37-45.

TitleCharacterization of hepcidin response to holotransferrin in novel recombinant TfR1 HepG2 cells
TypeJournal article
AuthorsMehta, K., Busbridge, Mark, Renshaw, D., Evans, R.W., Farnaud, S. and Patel, V.
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 (5 g/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 24 h and 48 h (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 4 h; 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 4 h of treatment, while HFE expression altered later at 24 h and 48 h, suggesting that TFR2 may function prior to HFE in HAMP regulation.

Keywordshepcidin
iron
HepG2
hepatocyte
transferrin
JournalBlood Cells, Molecules, and Diseases
Journal citation61, pp. 37-45
ISSN1079-9796
Year2016
PublisherElsevier
Publisher's version1-s2.0-S1079979616300808-main.pdf
Supplementary dataCharacterisation of hepcidin response manuscript suppl data.pdf
Digital Object Identifier (DOI)doi:10.1016/j.bcmd.2016.06.008
Publication dates
Published30 Jun 2016
LicenseCC BY-NC-ND 4.0

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Hunter, R.V., Patel, V., Baker, A.J. and Preedy, V.R. 2004. Liver dysfunction induced by bile duct ligation and galactosamine injection alters cardiac protein synthesis. Metabolism: Clinical and Experimental. 53 (8), pp. 964-968.

The plug domain of a neisserial TonB-dependent transporter retains structural integrity in the absence of its transmembrane β-barrel
Oke, M., Sarra, R., Ghirlando, R., Farnaud, S., Gorringe, A.R., Evans, R.W. and Buchanan, S.K. 2004. The plug domain of a neisserial TonB-dependent transporter retains structural integrity in the absence of its transmembrane β-barrel. FEBS Letters. 564 (3), pp. 294-300.

Interactions of lactoferricin-derived peptides with LPS and antimicrobial activity
Farnaud, S., Spiller, C., Moriarty, L.C., Patel, A., Gant, V., Odell, E.W. and Evans, R.W. 2004. Interactions of lactoferricin-derived peptides with LPS and antimicrobial activity. FEMS Microbiology Letters. 233 (2/29), pp. 193-199.

Lactoferrin: a multifunctional protein with antimicrobial properties
Farnaud, S. and Evans, R.W. 2003. Lactoferrin: a multifunctional protein with antimicrobial properties. Molecular Immunology. 40 (7), pp. 395-405.

Protein metabolism in alcohol misuse and toxicity
Preedy, V.R., Koll, M., Adachi, J., Mantle, D., Patel, V. and Peters, T. 2003. Protein metabolism in alcohol misuse and toxicity. in: Watson, R.R. and Preedy, V.R. (ed.) Nutrition and alcohol: linking nutrient interactions and dietary intake London CRC Press. pp. 261-300

Chronic ethanol consumption increases the concentration of a 4-hydroxynonenal adduct with the mitochondrial hydroxy methyl glutaryl CoA synthase in liver
Patel, V., Spencer, C. and Cunningham, C.C. 2003. Chronic ethanol consumption increases the concentration of a 4-hydroxynonenal adduct with the mitochondrial hydroxy methyl glutaryl CoA synthase in liver. Hepatology. 38, p. p393.

Emerging techniques in biomedical research and their application to alcohol toxicity
Patel, V., Chaurand, P., Caprioli, R., Austen, B., Frears, E., Manca, F., Davies, H., Vrana, K., Wheeler, M. and Preedy, V.R. 2003. Emerging techniques in biomedical research and their application to alcohol toxicity. Alcoholism: Clinical and Experimental Research. 27 (2), pp. 348-353.

Bacterial lipopolysaccharide directly stimulates cortisol secretion in human adrenal cells
Vakharia, K., Renshaw, D. and Hinson, J.P. 2002. Bacterial lipopolysaccharide directly stimulates cortisol secretion in human adrenal cells. Endocrine Research. 28 (4), pp. 357-361.

Bacterial Lipopolysaccharide directly stimulates cortisol secretion in human adrenal cells
Vakharia, K., Renshaw, D. and Hinson, J.P. 2002. Bacterial Lipopolysaccharide directly stimulates cortisol secretion in human adrenal cells. Endocrine Research. 28 (4), pp. 357-361.

Support for a three-dimensional structure predicting a Cys-Glu-Lys catalytic triad for Pseudomonas aeruginosa amidase comes from site-directed mutagenesis and mutations altering substrate specificity
Novo, C., Farnaud, S., Tata, R., Clemente, A. and Brown, P.R. 2002. Support for a three-dimensional structure predicting a Cys-Glu-Lys catalytic triad for Pseudomonas aeruginosa amidase comes from site-directed mutagenesis and mutations altering substrate specificity. Biochemical Journal. 365 (3), pp. 731-738.

Altered hepatic mitochondrial ribosome structure following chronic ethanol consumption
Patel, V. and Cunningham, C.C. 2002. Altered hepatic mitochondrial ribosome structure following chronic ethanol consumption. Archives of Biochemistry and Biophysics. 398 (1), pp. 41-50.

Chronic ethanol consumption increases the formation of an aldehyde-protein adduct in hepatic mitochondria
Patel, V., Young, T.A. and Cunningham, C.C. 2002. Chronic ethanol consumption increases the formation of an aldehyde-protein adduct in hepatic mitochondria. Alcoholism: Clinical and Experimental Research.

Regulation of rat adrenal vasoactive intestinal peptide content: effects of adrenocorticotropic hormone treatment and changes in dietary sodium intake
Hinson, J.P., Renshaw, D., Carroll, M. and Kapas, S. 2001. Regulation of rat adrenal vasoactive intestinal peptide content: effects of adrenocorticotropic hormone treatment and changes in dietary sodium intake. Journal of Neuroendocrinology. 13 (9), pp. 769-773.

Cardioprotective effect of propranolol from alcohol-induced heart muscle damage as assessed by plasma cardiac troponin-T.
Patel, V., Ajmal, R., Sherwood, R., Sullivan, A., Richardson, P. and Preedy, V.R. 2001. Cardioprotective effect of propranolol from alcohol-induced heart muscle damage as assessed by plasma cardiac troponin-T. Alcoholism: Clinical and Experimental Research. 25 (6), pp. 882-889.

Diarrhea reduces the rates of cardiac protein synthesis in myofibrillar protein fractions in rats in vivo
Hunter, R.V., Patel, V., Miell, J., Wong, H.J., Marway, J., Richardson, P. and Preedy, V.R. 2001. Diarrhea reduces the rates of cardiac protein synthesis in myofibrillar protein fractions in rats in vivo. Journal of Nutrition. 131 (5), pp. 1513-1519.

Acute doxorubicin (adriamycin) dosage does not reduce cardiac protein synthesis in vivo, but decreases diaminopeptidase I and proline endopeptidase activities
Zima, T., Tesar, V., Mantle, D., Koll, M., Patel, V., Richardson, P. and Preedy, V.R. 2001. Acute doxorubicin (adriamycin) dosage does not reduce cardiac protein synthesis in vivo, but decreases diaminopeptidase I and proline endopeptidase activities. Experimental and Molecular Pathology. 70 (2), pp. 154-161.

Physiochemical properties of rat liver mitochondrial ribosomes
Patel, V., Cunningham, C.C. and Hantgan, R. 2001. Physiochemical properties of rat liver mitochondrial ribosomes. Journal of Biological Chemistry. 276 (9), pp. 6739-6746.

In vivo protein synthetic rates of atrial, ventricular, and pulmonary tissue proteins in aortic constriction, goldblatt, and bromoethylamine models of hypertension
Siddiq, T., Patel, V., Sherwood, R., Richardson, P. and Preedy, V.R. 2001. In vivo protein synthetic rates of atrial, ventricular, and pulmonary tissue proteins in aortic constriction, goldblatt, and bromoethylamine models of hypertension. Experimental and Molecular Pathology. 70 (1), pp. 19-30.

Ethanol and oxidative stress
Sun, A., Ingelman-Sundberg, M., Neve, E., Matsumoto, H., Nishitani, Y., Minowa, Y., Fukui, Y., Bailey, S., Patel, V., Cunningham, C.C., Zima, T., Fialova, L., Mikulikova, L., Popov, P., Malbohan, I., Janebova, M., Nespor, K. and Sun, G. 2001. Ethanol and oxidative stress. Alcoholism: Clinical and Experimental Research. 25 (5 Suppl.), pp. 237S-243S.

Alpha-Tocopherol supplementation does not prevent acute alcohol-induced heart muscle damage
Koll, M., Patel, V., Sherwood, R.A., Seitz, H.K., Simanowski, U.A., Richardson, P.J., Peters, T.J. and Preedy, V.R. 2001. Alpha-Tocopherol supplementation does not prevent acute alcohol-induced heart muscle damage. Proceedings of the Nutrition Society. 60, p. P125A.

Ethanol and protein metabolism
Cunningham, C.C., Preedy, V.R., Paice, A., Hesketh, J., Peters, T., Patel, V., Volpi, E., Mawatari, K., Masaki, H., Mori, H. and Torii, K. 2001. Ethanol and protein metabolism. Alcoholism: Clinical and Experimental Research. 25 (5 Suppl.), pp. 262S-268S.

Chronic ethanol consumption alters the glutathione/glutathione peroxidase-1 system and protein oxidation status in rat liver
Bailey, S., Patel, V., Young, T.A., Asayama, K. and Cunningham, C.C. 2001. Chronic ethanol consumption alters the glutathione/glutathione peroxidase-1 system and protein oxidation status in rat liver. Alcoholism: Clinical and Experimental Research. 25 (5), pp. 726-733.

Evidence that cysteine-166 is the active-site nucleophile of Pseudomonas aeruginosa amidase: crystallization and preliminary X-ray diffraction analysis of the enzyme
Farnaud, S., Tata, R., Sohi, M.K., Wan, T., Brown, P.R. and Sutton, B.J. 1999. Evidence that cysteine-166 is the active-site nucleophile of Pseudomonas aeruginosa amidase: crystallization and preliminary X-ray diffraction analysis of the enzyme. Biochemical Journal. 340 (3), pp. 711-714.

Aluminium speciation: NMR studies of blood plasma, transferrin and citrate complexes
Bell, J.D., Evans, R.W., Kiang, W., Kubal, G., Radulovic, S., Sadler, P.J. and Williams, G. 1991. Aluminium speciation: NMR studies of blood plasma, transferrin and citrate complexes. Journal of Inorganic Biochemistry. 43 (2-3), p. 488.

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