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The effect of selective oestrogen receptor antagonists in an in vitro model of growth plate chondrogenesis

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Abstract

While oestrogen is recognized to play a key role in regulating growth, particularly in relation to epiphyseal fusion, the mechanisms that mediate its effects are still unclear. We utilized an in vitro model of chondrogenesis, the RCJ3.1C5.18 cell line, to explore the effect of oestrogen on this process. We demonstrated the presence of oestrogen receptors (ER) α and β in these cells, with increased abundance of both receptor sub-types evident as the cells differentiated. ERα localized to the nucleus, suggesting it was signalling by genomic pathways, while ERβ was seen predominantly in the cytoplasm, suggesting it may be utilizing non-genomic signalling. While exogenous oestrogen had no effect on proliferation or differentiation, we found some evidence for the endogenous production of oestrogen (intracrinology), as suggested by the expression of aromatase in these cells. Selective ERα blockade with methyl piperidinopyrazole (MPP) led to a significant reduction in both proliferation and differentiation, while ERβ blockade with R,R tetrahydrochrysene (THC) led to an increase in these parameters. This is in keeping with results from mouse knockout models suggesting that unopposed ERβ signalling leads to an inhibition of skeletal growth. Our results are further evidence for the importance of differential ER signalling in regulating chondrogenesis. Future studies examining in vivo effects of these agents are required to extrapolate these findings to a mammalian model.

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Acknowledgments

We would like to thank Sandra Higgins for her assistance, particularly with the real-time PCR, as well as Walid Azar for undertaking the semi-quantification of the immunocytochemistry slides. Dr Simm was the recipient of a Pfizer Australian Paediatric Endocrine Care Grant that allowed him to complete this research.

Disclosure

Dr Simm was supported by an Australian Paediatric Endocrine Care Grant (Pfizer) during the completion of this work. Prof Werther and Dr Russo have nothing to declare in relation to this manuscript.

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Authors and Affiliations

  1. Department of Endocrinology and Diabetes, Royal Children’s Hospital Melbourne, Flemington Rd, Parkville, VIC, 3052, Australia

    Peter J. Simm & George A. Werther

  2. Murdoch Childrens Research Institute, Centre for Hormone Research, Parkville, Australia

    Peter J. Simm, Vincenzo C. Russo & George A. Werther

  3. Department of Paediatrics, University of Melbourne, Parkville, Australia

    Peter J. Simm, Vincenzo C. Russo & George A. Werther

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  1. Peter J. Simm
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  2. Vincenzo C. Russo
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  3. George A. Werther
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Correspondence to Peter J. Simm.

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Simm, P.J., Russo, V.C. & Werther, G.A. The effect of selective oestrogen receptor antagonists in an in vitro model of growth plate chondrogenesis. Endocrine 40, 27–34 (2011). https://doi.org/10.1007/s12020-011-9473-2

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  • DOI: https://doi.org/10.1007/s12020-011-9473-2

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