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Stimulation of neuroendocrine differentiation in prostate cancer cells by GHRH and its blockade by GHRH antagonists

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Summary

Prostate cancer is the second leading cause of cancer-related deaths among men in developed countries. Neuroendocrine prostate cancer, in particular, is associated with an aggressive phenotype and a poor prognosis. Neuroendocrine cells produce and secrete peptide hormones and growth factors in a paracrine/autocrine manner which promote the progression of the disease. Recent studies have demonstrated that extracellular vesicles or exosomes are released by prostate cancer cells, supporting the spread of prostate cancer. Hence, the aim of this study was to investigate the effect of growth hormone-releasing hormone (GHRH) on neuroendocrine differentiation (NED) in the androgen-dependent prostate cancer cell line LNCaP and the molecular mechanisms underlying these effects. GHRH induced an increase in the percentage of neurite-bearing cells and in the protein levels of Neuron-Specific Enolase. Both effects were blocked by the GHRH receptor antagonist MIA-690. In addition, pretreatment of these cells with the calcium chelator BAPTA, the EGFR inhibitor AG-1478 or the HER2 inhibitor AG-825 reduced the effect of GHRH, suggesting that the GHRH-induced stimulation of NED involves calcium channel activation and EGFR/HER2 transactivation. Finally, PC3-derived exosomes led to an increase in NED, cell proliferation and cell adhesion. Altogether, these findings suggest that GHRH antagonists should be considered for in the management of neuroendocrine prostate cancer.

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Abbreviations

PCa:

Prostate cancer

CRPCa:

castration resistant prostate cancer

NEPCa:

neuroendocrine prostate cancer

NE:

Neuroendocrine

NED:

neuroendocrine differentiation

GHRH:

growth hormone-releasing hormone

GHRH-R:

growth hormone-releasing hormone receptors

VIP:

vasoactive intestinal peptide

EVs:

extracellular vesicles

FBS:

fetal bovine serum

BrdU:

bromodeoxyuridine

NSE:

neuron-specific enolase

PBS:

phosphate buffered saline

EDTA:

ethylenediaminetetraacetic acid

SE:

standard error

PKA:

protein kinase A

ERK:

extracellular signal-regulated kinases

MAPK:

mitogen-activated protein kinase

MEK:

mitogen-activated protein kinase kinase

PI3K:

phosphoinositide 3-kinase

EGFR:

epidermal growth factor receptor

HER2:

human epidermal growth factor receptor 2

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Acknowledgments

We thank Dr. Puebla (University of Alcala) for her assistance in writing the manuscript.

Funding

This work was supported by grants from Universidad de Alcalá (CCG2015/BIO-010 to A.M.B. and CCG2014/BIO-028 to A.M.B.)

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Authors and Affiliations

  1. Department of Systems Biology, University of Alcalá, 28871, Alcalá de Henares, Spain

    Laura Muñoz-Moreno, María J. Carmena, Juan C. Prieto & Ana M. Bajo

  2. Veterans Affairs Medical Center, Endocrine, Polypeptide and Cancer Institute, Miami, FL, 33125, USA

    Andrew V. Schally

  3. Departments of Pathology and Medicine, Division of Medical Oncology, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA

    Andrew V. Schally

  4. Sylvester Comprehensive Cancer Center, Miller School of Medicine, University of Miami, Miami, FL, 33136, USA

    Andrew V. Schally

Authors
  1. Laura Muñoz-Moreno
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  2. María J. Carmena
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  3. Andrew V. Schally
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  5. Ana M. Bajo
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Correspondence to Ana M. Bajo.

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The author Laura Muñoz-Moreno declares that she has no conflict of interest. The author María J. Carmena declares that she has no conflict of interest. The author Andrew V. Schally declares that he has no conflict of interest. The author Juan C. Prieto declares that he has no conflict of interest. The author Ana M. Bajo declares that she has no conflict of interest.

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Muñoz-Moreno, L., Carmena, M.J., Schally, A.V. et al. Stimulation of neuroendocrine differentiation in prostate cancer cells by GHRH and its blockade by GHRH antagonists. Invest New Drugs 38, 746–754 (2020). https://doi.org/10.1007/s10637-019-00831-2

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