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Ghrelin and des-acyl ghrelin inhibit aromatase expression and activity in human adipose stromal cells: suppression of cAMP as a possible mechanism

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Abstract

Aromatase converts androgens into estrogens and its expression within adipose stromal cells (ASCs) is believed to be the major driver of estrogen-dependent cancers in older women. Ghrelin is a gut-hormone that is involved in the regulation of appetite and known to bind to and activate the cognate ghrelin receptor, GHSR1a. The unacylated form of ghrelin, des-acyl ghrelin, binds weakly to GHSR1a but has been shown to play an important role in regulating a number of physiological processes. The aim of this study was to determine the effect of ghrelin and des-acyl ghrelin on aromatase in primary human ASCs. Primary human ASCs were isolated from adipose tissue of women undergoing cosmetic surgery. Real-time PCR and tritiated water-release assays were performed to examine the effect of treatment on aromatase transcript expression and aromatase activity, respectively. Treatments included ghrelin, des-acyl ghrelin, obestatin, and capromorelin (GHSR1a agonist). GHSR1a protein expression was assessed by Western blot and effects of treatment on Ca2+ and cAMP second messenger systems were examined using the Flexstation assay and the Lance Ultra cAMP kit, respectively. Results demonstrate that pM concentrations of ghrelin and des-acyl ghrelin inhibit aromatase transcript expression and activity in ASCs under basal conditions and in PGE2-stimulated cells. Moreover, the effects of ghrelin and des-acyl ghrelin are mediated via effects on aromatase promoter PII-specific transcripts. Neither the GHSR1a-specific agonist capromorelin nor obestatin had any effect on aromatase transcript expression or activity. Moreover, GHSR1a protein was undetectable by Western blot and neither ghrelin nor capromorelin elicited a calcium response in ASCs. Finally, ghrelin caused a significant decrease in basal and forskolin-stimulated cAMP in ASC. These findings suggest that ghrelin acts at alternate receptors in ASCs by decreasing intracellular cAMP levels. Ghrelin mimetics may be useful in the treatment of estrogen-dependent breast cancer.

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Acknowledgments

This research was supported by NHMRC (Australia) Project Grant # GNT1005735 and by a grant from the National Breast Cancer Foundation (NC-14-011) to KAB, and by the Victorian Government Operational Infrastructure Support Program. KAB is supported by an NHMRC (Australia) Career Development Award GNT1007714. We thank Mr Seungmin Ham for his technical support.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

The experiments detailed in the present manuscript were performed in accordance with National Health and Medical Research Council of Australia (NHMR) guidelines.

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Authors and Affiliations

  1. Metabolism & Cancer Laboratory, MIMR-PHI Institute of Medical Research, Clayton, VIC, 3168, Australia

    Maria M. Docanto, Fangyuan Yang, CheukMan C. Au, Rahini Ragavan, Xuyi Wang & Kristy A. Brown

  2. Department of Anatomy & Neuroscience, University of Melbourne, Parkville, VIC, 3010, Australia

    Brid Callaghan & John B. Furness

  3. Monash Institute of Medical Research, Monash University, Clayton, VIC, 3168, Australia

    CheukMan C. Au & Kristy A. Brown

  4. Department of Physiology, Monash University, Clayton, VIC, 3168, Australia

    Rahini Ragavan, Xuyi Wang, Zane B. Andrews & Kristy A. Brown

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  1. Maria M. Docanto
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  2. Fangyuan Yang
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  3. Brid Callaghan
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Correspondence to Kristy A. Brown.

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Docanto, M.M., Yang, F., Callaghan, B. et al. Ghrelin and des-acyl ghrelin inhibit aromatase expression and activity in human adipose stromal cells: suppression of cAMP as a possible mechanism. Breast Cancer Res Treat 147, 193–201 (2014). https://doi.org/10.1007/s10549-014-3060-1

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