Characterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells

Mehta, K., Greenwell, P., Renshaw, D., Busbridge, Mark, Garcia, M, Farnaud, S. and Patel, V. B. 2015. Characterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells. Blood Cells, Molecules, and Diseases. 55 (2), pp. 110-118.

TitleCharacterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells
AuthorsMehta, K., Greenwell, P., Renshaw, D., Busbridge, Mark, Garcia, M, Farnaud, S. and Patel, V. B.
Abstract

Iron overload coupled with low hepcidin levels are characteristics of hereditary haemochromatosis. To understand the role of transferrin receptor (TFR) and intracellular iron in hepcidin secretion, Chinese hamster ovary transferrin receptor variant (CHO TRVb-1) cells were used that express iron-response-element-depleted human TFRC mRNA (TFRC∆IRE). Results showed that CHO TRVb-1 cells expressed higher basal levels of cell-surface TFR1 than HepG2 cells (2.2-fold; p < 0.01) and following 5 g/L holotransferrin treatment maintained constitutive over-expression at 24h and 48 h, contrasting the HepG2 cells where the receptor levels significantly declined. Despite this, the intracellular iron content was neither higher than HepG2 cells nor increased over time under basal or holotransferrin-treated conditions. Interestingly, hepcidin secretion in CHO TRVb-1 cells exceeded basal levels at all time-points (p < 0.02) and matched levels in HepG2 cells following treatment. While TFRC mRNA expression showed expected elevation (2h, p < 0.03; 4h; p < 0.05), slc40a1 mRNA expression was also elevated (2 h, p < 0.05; 4 h, p < 0.03), unlike the HepG2 cells. In conclusion, the CHO TRVb-1 cells prevented cellular iron-overload by elevating slc40a1 expression, thereby highlighting its significance in the absence of iron-regulated TFRC mRNA. Furthermore, hepcidin response to holotransferrin treatment was similar to HepG2 cells and resembled the human physiological response.

KeywordsHepcidin; Iron regulation; Iron response element; Transferrin receptor
JournalBlood Cells, Molecules, and Diseases
Journal citation55 (2), pp. 110-118
ISSN1079-9796
Year2015
PublisherElsevier
Accepted author manuscriptCharacterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells.pdf
Digital Object Identifier (DOI)doi:10.1016/j.bcmd.2015.05.002
Publication dates
Published08 May 2015
LicenseCC BY-NC-ND 4.0

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Rughooputh, S., Manraj, S., Eddoo, R. and Greenwell, P. 2009. Expression of the c-myc oncogene and the presence of HPV 18: possible surrogate markers for cervical cancer? British Journal of Biomedical Science. 66 (2), pp. 74-78.

ABO reverse grouping: effect of varying concentrations of the enzyme bromelain
Rookard, L.E., Edmondson, O. and Greenwell, P. 2009. ABO reverse grouping: effect of varying concentrations of the enzyme bromelain. British Journal of Biomedical Science. 66 (2), pp. 93-97.

Liver metabolism: biochemical and molecular regulations
Gyamfi, D. and Patel, V. 2009. Liver metabolism: biochemical and molecular regulations. in: Preedy, V.R. (ed.) Nutrition, diet therapy, and the liver London CRC Press. pp. 3-15

Hepatic oxidative stress and apoptosis following acute alcohol
Everitt, H.E., Tewfik, I., Preedy, V.R. and Patel, V. 2009. Hepatic oxidative stress and apoptosis following acute alcohol. Alcohol & Alcoholism. 44, p. p85.

Annexin 1 mediates the rapid anti-inflammatory effects of neutrophil-derived microparticles
Dalli, J., Norling, L.V., Renshaw, D., Cooper, D., Leung Kit-Y and Perretti, M. 2008. Annexin 1 mediates the rapid anti-inflammatory effects of neutrophil-derived microparticles. Blood. 112 (6), pp. 2512-2519.

Purification and analysis of DNases of Tritrichomonas foetus: evidence that these enzymes are glycoproteins
Greenwell, P., Younes, M. and Rughooputh, S. 2008. Purification and analysis of DNases of Tritrichomonas foetus: evidence that these enzymes are glycoproteins. International Journal for Parasitology. 38 (7), pp. 749-756.

Tomatoes: origin, cultivation techniques, and germplasm resources
da Silva, D.J.H., Abreu, F.B., Caliman, F.R.B., Antonio, A.C. and Patel, V. 2008. Tomatoes: origin, cultivation techniques, and germplasm resources. in: Preedy, V.R. and Watson, R.R. (ed.) Tomatoes and tomato products: nutritional, medicinal and therapeutic properties CRC Press. pp. 3-26

Effects of 4-hydroxynonenal on mitochondrial 3-hydroxy-3methylglutaryl (HMG-CoA) synthase
Patel, V., Spencer, C.H., Young, T.A., Lively, M.O. and Cunningham, C.C. 2007. Effects of 4-hydroxynonenal on mitochondrial 3-hydroxy-3methylglutaryl (HMG-CoA) synthase. Free Radical Biology and Medicine. 43 (11), pp. 1499-1507.

Proteome analysis of metastatic colorectal cancer cells recognized by the lectin Helix pomatia agglutinin (HPA)
Saint-Guirons, J., Zeqiraj, E., Schumacher, U., Greenwell, P. and Dwek, M. 2007. Proteome analysis of metastatic colorectal cancer cells recognized by the lectin Helix pomatia agglutinin (HPA). Proteomics. 7 (22), pp. 4082-4089.

Therapeutic exploitation of endogenous anti-inflammatory mechanisms: old and new leads
Renshaw, D., Urquhart, P. and Perretti, M. 2007. Therapeutic exploitation of endogenous anti-inflammatory mechanisms: old and new leads. Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry. 5 (4), pp. 289-305.

Identification and elimination of false-positives in an ELISA-based system for qualitative assessment of glycoconjugate binding using a selection of plant lectins
Afrough, B., Dwek, M. and Greenwell, P. 2007. Identification and elimination of false-positives in an ELISA-based system for qualitative assessment of glycoconjugate binding using a selection of plant lectins. BioTechniques. 43 (4), pp. 458-462.

Chikungunya virus: an emerging problem?
Rughooputh, S. and Greenwell, P. 2007. Chikungunya virus: an emerging problem? Biomedical Scientist.

Nutrition and alcoholic liver disease
Everitt, H.E., Patel, V. and Tewfik, I. 2007. Nutrition and alcoholic liver disease. Nutrition Bulletin. 32 (2), pp. 138-144.

Prevalence of human papillomavirus amongst cervical cancer sufferers amongst slum dwellers in India
Rughooputh, S., Kachaliya, S., Jetly, D. and Greenwell, P. 2007. Prevalence of human papillomavirus amongst cervical cancer sufferers amongst slum dwellers in India. British Journal of Biomedical Science. 64 (1), pp. 28-31.

Cervical cancer and human papillomavirus among slum dwellers in India
Rughooputh, S., Kachaliya, S., Jetly, D. and Greenwell, P. 2007. Cervical cancer and human papillomavirus among slum dwellers in India. British Journal of Biomedical Science. 64 (1), pp. 28-31.

Adrenomedullin increases the expression of calcitonin-like receptor and receptor activity modifying protein 2 mRNA in human microvascular endothelial cells
Schwarz, N., Renshaw, D., Kapas, S. and Hinson, J.P. 2006. Adrenomedullin increases the expression of calcitonin-like receptor and receptor activity modifying protein 2 mRNA in human microvascular endothelial cells. Journal of Endocrinology. 190 (2), pp. 505-514.

Intra-adrenal mechanisms in the response to chronic stress: investigation in a rat model of emotionality
Kosti, O., Raven, P.W., Renshaw, D. and Hinson, J.P. 2006. Intra-adrenal mechanisms in the response to chronic stress: investigation in a rat model of emotionality. Journal of Endocrinology. 189 (2), pp. 211-218.

Detection of human papillomavirus from archival tissues in cervical cancer patients in Mauritius
Rughooputh, S., Eddoo, R., Manraj, S., Jeebun, N. and Greenwell, P. 2006. Detection of human papillomavirus from archival tissues in cervical cancer patients in Mauritius. Journal of Clinical Virology. 35 (2), pp. 173-178.

A proteomic approach to identify the integrins α 6 and α V as HPA binding partners in the metastatic colorectal cancer cell line HT29
Saint-Guirons, J., Zeqiraj, E., Greenwell, P. and Dwek, M. 2006. A proteomic approach to identify the integrins α 6 and α V as HPA binding partners in the metastatic colorectal cancer cell line HT29. Molecular and Cellular Proteomics. 5 (10 (supplement)), p. S148.

Association between HPV and C-MYC gene expression in cervical cancer
Rughooputh, S., Eddoo, R., Manraj, S., Jeebun, N. and Greenwell, P. 2006. Association between HPV and C-MYC gene expression in cervical cancer. International Journal of Molecular Medicine. 18 (supplement).

Analysis of glycosidases activity in Chlamydia trachomatis L2 serotype
Greenwell, P., Kakourou, G. and Rughooputh, S. 2006. Analysis of glycosidases activity in Chlamydia trachomatis L2 serotype. Internet Journal of Medical Update. 1 (1).

Protein adduct species in muscle and liver of rats following acute ethanol administration
Patel, V., Worrall, S., Emery, P.W. and Preedy, V.R. 2005. Protein adduct species in muscle and liver of rats following acute ethanol administration. Alcohol & Alcoholism. 40 (6), pp. 485-493.

Vaccination against cervical cancer
Greenwell, P. and Rughooputh, S. 2005. Vaccination against cervical cancer. Biomedical Scientist. 49 (6), p. 441.

Proteomics: more than just biochemistry
Greenwell, P. and Rughooputh, S. 2005. Proteomics: more than just biochemistry. Biomedical Scientist. 49 (6), pp. 459-460.

Trichomonas vaginalis: paradigm of a successful sexually transmitted organism
Rughooputh, S. and Greenwell, P. 2005. Trichomonas vaginalis: paradigm of a successful sexually transmitted organism. British Journal of Biomedical Science. 62 (4), pp. 193-200.

Application of recombinant phage display antibody system in study of Codakia orbicularis gill proteins
Gourdine, J.P., Greenwell, P. and Smith-Ravin, E.J. 2005. Application of recombinant phage display antibody system in study of Codakia orbicularis gill proteins. Applied Biochemistry and Biotechnology. 125 (1), pp. 41-52.

The Unani system of medicine: does it have a scientific basis?
Syeda, A., Rughooputh, S. and Greenwell, P. 2004. The Unani system of medicine: does it have a scientific basis? Biomedical Scientist. 48 (9), pp. 971-972.

Liver dysfunction induced by bile duct ligation and galactosamine injection alters cardiac protein synthesis
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.

Genetically modified food: good news but bad press
Greenwell, P. and Rughooputh, S. 2004. Genetically modified food: good news but bad press. Biomedical Scientist. 48 (8), pp. 845-846.

Bioinformatics: a useful tool for the molecular microbiologist?
Greenwell, P. and Rughooputh, S. 2004. Bioinformatics: a useful tool for the molecular microbiologist? Biomedical Scientist. 48 (5), pp. 494-495.

Detection of human papillomavirus from liquid-based cytology specimens by in-house PCR: a pilot study
Rughooputh, S., Parmar, K. and Greenwell, P. 2004. Detection of human papillomavirus from liquid-based cytology specimens by in-house PCR: a pilot study. British Journal of Biomedical Science. 61 (1), pp. 22-25.

Use of bioinformatics and PCR in the search for ABC transporter homology among various bacteria
Jauhangeer, B.R., Wren, M.W.D., MacDonald, R.A.C., Perry, D. and Greenwell, P. 2004. Use of bioinformatics and PCR in the search for ABC transporter homology among various bacteria. British Journal of Biomedical Science. 61 (4), pp. 182-185.

Trichomonas vaginalis: an irritating protozoan or an important viral co-factor
Greenwell, P. and Rughooputh, S. 2004. Trichomonas vaginalis: an irritating protozoan or an important viral co-factor. Biomedical Scientist. 48 (1), pp. 40-41.

Human papillomavirus: an overview
Rughooputh, S. and Greenwell, P. 2003. Human papillomavirus: an overview. Biomedical Scientist. 47, pp. 1070-1071.

Isolation of Neisseria gonorrhoeae-specific antibodies from a phage display library
Rughooputh, S. and Greenwell, P. 2003. Isolation of Neisseria gonorrhoeae-specific antibodies from a phage display library. Molecular and Cellular Proteomics. 2 (9), p. 758.

Bioinformatics in the identification of glycosidase genes in the Chlamydia genome
Greenwell, P. and Rughooputh, S. 2003. Bioinformatics in the identification of glycosidase genes in the Chlamydia genome. Molecular and Cellular Proteomics. 2 (9), p. 826.

Phage display antibody against gill proteins of the tropical clam
Gourdine, J.P., Smith-Ravin, E.J. and Greenwell, P. 2003. Phage display antibody against gill proteins of the tropical clam. Molecular and Cellular Proteomics. 2 (9), p. 759.

Incidence and prognostic significance of C-MPL expression in acute myeloid leukemia
Chelvatheebam, S., Langabeer, S., Hills, R., Greenwell, P. and Linch, D. 2003. Incidence and prognostic significance of C-MPL expression in acute myeloid leukemia. Leukemia Research. 27 (9), pp. 839-870.

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.

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.

Beyond hybridoma technology: production of monoclonal antibodies by phage display
Greenwell, P. and Rughooputh, S. 2001. Beyond hybridoma technology: production of monoclonal antibodies by phage display. Biomedical Scientist. 45, pp. 574-575.

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.

Isoforms of Na+, K+-ATPase in human prostate; specificity of expression and apical membrane polarization
Mobasheri, A., Oukrif, D., Dawodu, S.P., Sinha, M., Greenwell, P., Stewart, D., Djamgoz, M.B.A., Foster, C.S., Martin-Vasallo, P. and Mobasheri, R. 2001. Isoforms of Na+, K+-ATPase in human prostate; specificity of expression and apical membrane polarization. Histology and Histopathology. 16 (1), pp. 141-154.

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.

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