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01-10-2012 | Original Article

Iron excess limits HHIPL-2 gene expression and decreases osteoblastic activity in human MG-63 cells

Authors: M. Doyard, N. Fatih, A. Monnier, M. L. Island, M. Aubry, P. Leroyer, R. Bouvet, G. Chalès, J. Mosser, O. Loréal, P. Guggenbuhl

Published in: Osteoporosis International | Issue 10/2012

Abstract

Summary

In order to understand mechanisms involved in osteoporosis observed during iron overload diseases, we analyzed the impact of iron on a human osteoblast-like cell line. Iron exposure decreases osteoblast phenotype. HHIPL-2 is an iron-modulated gene which could contribute to these alterations. Our results suggest osteoblast impairment in iron-related osteoporosis.

Introduction

Iron overload may cause osteoporosis. An iron-related decrease in osteoblast activity has been suggested.

Methods

We investigated the effect of iron exposure on human osteoblast cells (MG-63) by analyzing the impact of ferric ammonium citrate (FAC) and iron citrate (FeCi) on the expression of genes involved in iron metabolism or associated with osteoblast phenotype. A transcriptomic analysis was performed to identify iron-modulated genes.

Results

FAC and FeCi exposure modulated cellular iron status with a decrease in TFRC mRNA level and an increase in intracellular ferritin level. FAC increased ROS level and caspase 3 activity. Ferroportin, HFE and TFR2 mRNAs were expressed in MG-63 cells under basal conditions. The level of ferroportin mRNA was increased by iron, whereas HFE mRNA level was decreased. The level of mRNA alpha 1 collagen type I chain, osteocalcin and the transcriptional factor RUNX2 were decreased by iron. Transcriptomic analysis revealed that the mRNA level of HedgeHog Interacting Protein Like-2 (HHIPL-2) gene, encoding an inhibitor of the hedgehog signaling pathway, was decreased in the presence of FAC. Specific inhibition of HHIPL-2 expression decreased osteoblast marker mRNA levels. Purmorphamine, hedgehog pathway activator, increased the mRNA level of GLI1, a target gene for the hedgehog pathway, and decreased osteoblast marker levels. GLI1 mRNA level was increased under iron exposure.

Conclusion

We showed that in human MG-63 cells, iron exposure impacts iron metabolism and osteoblast gene expression. HHIPL-2 gene expression modulation may contribute to these alterations. Our results support a role of osteoblast impairment in iron-related osteoporosis.

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Title: Iron excess limits HHIPL-2 gene expression and decreases osteoblastic activity in human MG-63 cells
Authors: M. Doyard
N. Fatih
A. Monnier
M. L. Island
M. Aubry
P. Leroyer
R. Bouvet
G. Chalès
J. Mosser
O. Loréal
P. Guggenbuhl
Publication date: 01-10-2012
Publisher: Springer-Verlag
Published in: Osteoporosis International / Issue 10/2012
Print ISSN: 0937-941X
Electronic ISSN: 1433-2965
DOI: https://doi.org/10.1007/s00198-011-1871-z

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Iron excess limits HHIPL-2 gene expression and decreases osteoblastic activity in human MG-63 cells

Abstract

Summary

Introduction

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Summary

Introduction

Methods

Results

Conclusion

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