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Int. J. Mol. Sci. 2017, 18(5), 926; doi:10.3390/ijms18050926

Estrogen Modulates Specific Life and Death Signals Induced by LH and hCG in Human Primary Granulosa Cells In Vitro

1
Unit of Endocrinology, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, NOCSAE, via P. Giardini 1355, 41126 Modena, Italy
2
Center for Genomic Research, University of Modena and Reggio Emilia, Via G. Campi 287, 41125 Modena, Italy
3
Department of Medicine, Endocrinology, Metabolism and Geriatrics, Azienda USL di Modena, 41124 Modena, Italy
4
Units of Obstetrics and Gynecology, IRCCS-Arcispedale Santa Maria Nuova, Via Risorgimento 80, 42123 Reggio Emilia, Italy
5
Department of Medical and Surgical Sciences for Children and Adults, University of Modena and Reggio Emilia, Via del Pozzo 71, 41124 Modena, Italy
*
Author to whom correspondence should be addressed.
Academic Editor: Udo Jeschke
Received: 28 February 2017 / Revised: 31 March 2017 / Accepted: 25 April 2017 / Published: 28 April 2017
(This article belongs to the Special Issue hCG—An Endocrine, Regulator of Gestation and Cancer)
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Abstract

Luteinizing hormone (LH) and human chorionic gonadotropin (hCG) are glycoprotein hormones used for assisted reproduction acting on the same receptor (LHCGR) and mediating different intracellular signaling. We evaluated the pro- and anti-apoptotic effect of 100 pM LH or hCG, in the presence or in the absence of 200 pg/mL 17β-estradiol, in long-term, serum-starved human primary granulosa cells (hGLC) and a transfected granulosa cell line overexpressing LHCGR (hGL5/LHCGR). To this purpose, phospho-extracellular-regulated kinase 1/2 (pERK1/2), protein kinase B (pAKT), cAMP-responsive element binding protein (pCREB) activation and procaspase 3 cleavage were evaluated over three days by Western blotting, along with the expression of target genes by real-time PCR and cell viability by colorimetric assay. We found that LH induced predominant pERK1/2 and pAKT activation STARD1, CCND2 and anti-apoptotic XIAP gene expression, while hCG mediated more potent CREB phosphorylation, expression of CYP19A1 and procaspase 3 cleavage than LH. Cell treatment by LH is accompanied by increased (serum-starved) cell viability, while hCG decreased the number of viable cells. The hCG-specific, pro-apoptotic effect was blocked by a physiological dose of 17β-estradiol, resulting in pAKT activation, lack of procaspase 3 cleavage and increased cell viability. These results confirm that relatively high levels of steroidogenic pathway activation are linked to pro-apoptotic signals in vitro, which may be counteracted by other factors, i.e., estrogens. View Full-Text
Keywords: LH; hCG; gonadotropins; apoptosis; granulosa LH; hCG; gonadotropins; apoptosis; granulosa
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MDPI and ACS Style

Casarini, L.; Riccetti, L.; De Pascali, F.; Gilioli, L.; Marino, M.; Vecchi, E.; Morini, D.; Nicoli, A.; La Sala, G.B.; Simoni, M. Estrogen Modulates Specific Life and Death Signals Induced by LH and hCG in Human Primary Granulosa Cells In Vitro. Int. J. Mol. Sci. 2017, 18, 926.

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