Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells

Iron is essential for all living organisms. It is strictly controlled by iron transporters, transferrin receptors, ferroportin and hepcidin. Erythroferrone (ERFE) is an iron-regulatory hormone which is highly expressed in erythroblasts by erythropoietin (EPO) stimulation and osteoblasts independentl...

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Autores principales:	Min Min Than, Pimpisid Koonyosying, Jetsada Ruangsuriya, Sunhawit Junrungsee, Chairat Uthaipibull, Somdet Srichairatanakool
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	erythroferrone hepcidin iron recombinant protein receptor Technology T Electrical engineering. Electronics. Nuclear engineering TK1-9971 Engineering (General). Civil engineering (General) TA1-2040 Microscopy QH201-278.5 Descriptive and experimental mechanics QC120-168.85
Acceso en línea:	https://doaj.org/article/676b77ba2ec047c883ce228698ab1928
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spelling	oai:doaj.org-article:676b77ba2ec047c883ce228698ab19282021-11-11T18:03:20ZEffect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells10.3390/ma142164801996-1944https://doaj.org/article/676b77ba2ec047c883ce228698ab19282021-10-01T00:00:00Zhttps://www.mdpi.com/1996-1944/14/21/6480https://doaj.org/toc/1996-1944Iron is essential for all living organisms. It is strictly controlled by iron transporters, transferrin receptors, ferroportin and hepcidin. Erythroferrone (ERFE) is an iron-regulatory hormone which is highly expressed in erythroblasts by erythropoietin (EPO) stimulation and osteoblasts independently of EPO by sequestering bone morphogenetic proteins and inhibiting hepatic hepcidin expression. Although the hepcidin suppressive function of ERFE is known, its receptors still require investigation. Here, we aim to identify ERFE receptors on the HepG2 and Huh7 cells responsible for ERFE. Recombinant ERFE (rERFE) was first produced in HEK293 cells transfected with pcDNA3.1 + ERFE, then purified and detected by Western blot. The liver cells were treated with an rERFE-rich medium of transfected HEK293 cells and a purified rERFE-supplemented medium at various time points, and hepcidin gene (<i>Hamp1</i>) expression was determined using qRT-PCR. The results show that 37-kD rERFE was expressed in HEK293 cells. <i>Hamp1</i> was suppressed at 3 h and 6 h in Huh7 cells after rERFE treatments (<i>p</i> < 0.05), then restored to the original levels. <i>Hamp1</i> was activated after treatment with purified rERFE for 24 h and 48 h. Together, these results reveal that ERFE suppressed <i>Hamp1</i> expression in liver cells, possibly acting on membrane ERFE receptor, which in Huh7 cells was more sensitive to the ERFE concentrate.Min Min ThanPimpisid KoonyosyingJetsada RuangsuriyaSunhawit JunrungseeChairat UthaipibullSomdet SrichairatanakoolMDPI AGarticleerythroferronehepcidinironrecombinant proteinreceptorTechnologyTElectrical engineering. Electronics. Nuclear engineeringTK1-9971Engineering (General). Civil engineering (General)TA1-2040MicroscopyQH201-278.5Descriptive and experimental mechanicsQC120-168.85ENMaterials, Vol 14, Iss 6480, p 6480 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	erythroferrone hepcidin iron recombinant protein receptor Technology T Electrical engineering. Electronics. Nuclear engineering TK1-9971 Engineering (General). Civil engineering (General) TA1-2040 Microscopy QH201-278.5 Descriptive and experimental mechanics QC120-168.85
spellingShingle	erythroferrone hepcidin iron recombinant protein receptor Technology T Electrical engineering. Electronics. Nuclear engineering TK1-9971 Engineering (General). Civil engineering (General) TA1-2040 Microscopy QH201-278.5 Descriptive and experimental mechanics QC120-168.85 Min Min Than Pimpisid Koonyosying Jetsada Ruangsuriya Sunhawit Junrungsee Chairat Uthaipibull Somdet Srichairatanakool Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells
description	Iron is essential for all living organisms. It is strictly controlled by iron transporters, transferrin receptors, ferroportin and hepcidin. Erythroferrone (ERFE) is an iron-regulatory hormone which is highly expressed in erythroblasts by erythropoietin (EPO) stimulation and osteoblasts independently of EPO by sequestering bone morphogenetic proteins and inhibiting hepatic hepcidin expression. Although the hepcidin suppressive function of ERFE is known, its receptors still require investigation. Here, we aim to identify ERFE receptors on the HepG2 and Huh7 cells responsible for ERFE. Recombinant ERFE (rERFE) was first produced in HEK293 cells transfected with pcDNA3.1 + ERFE, then purified and detected by Western blot. The liver cells were treated with an rERFE-rich medium of transfected HEK293 cells and a purified rERFE-supplemented medium at various time points, and hepcidin gene (<i>Hamp1</i>) expression was determined using qRT-PCR. The results show that 37-kD rERFE was expressed in HEK293 cells. <i>Hamp1</i> was suppressed at 3 h and 6 h in Huh7 cells after rERFE treatments (<i>p</i> < 0.05), then restored to the original levels. <i>Hamp1</i> was activated after treatment with purified rERFE for 24 h and 48 h. Together, these results reveal that ERFE suppressed <i>Hamp1</i> expression in liver cells, possibly acting on membrane ERFE receptor, which in Huh7 cells was more sensitive to the ERFE concentrate.
format	article
author	Min Min Than Pimpisid Koonyosying Jetsada Ruangsuriya Sunhawit Junrungsee Chairat Uthaipibull Somdet Srichairatanakool
author_facet	Min Min Than Pimpisid Koonyosying Jetsada Ruangsuriya Sunhawit Junrungsee Chairat Uthaipibull Somdet Srichairatanakool
author_sort	Min Min Than
title	Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells
title_short	Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells
title_full	Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells
title_fullStr	Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells
title_full_unstemmed	Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells
title_sort	effect of recombinant human erythroferrone protein on hepcidin gene (<i>hamp1</i>) expression in hepg2 and huh7 cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/676b77ba2ec047c883ce228698ab1928
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Effect of Recombinant Human Erythroferrone Protein on Hepcidin Gene (<i>Hamp1</i>) Expression in HepG2 and HuH7 Cells

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