scholarly journals Neuromedin B Receptor Antagonist Suppresses Pomc Expression in AtT-20 Cells and Human Corticotroph Adenoma Cells

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A546-A547
Author(s):  
Tomonori Sekizaki ◽  
Hiraku Kameda ◽  
Akinobu Nakamura ◽  
Hiroshi Nomoto ◽  
Kyu Yong Cho ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Pituitary Adenoma ◽  
Mrna Expression ◽  
Real Time ◽  
Hormone Secretion ◽  
Corticotroph Adenoma ◽  
Adenoma Cell ◽  
Pomc Mrna ◽  
Post Hoc ◽  
Real Time Qpcr

Abstract Objective: We previously reported that Neuromedin B (NMB) is expressed in murine pituitary corticotrophs under adrenal insufficiency (1). Because NMB is also expressed in several cancer cells and stimulates ACTH secretion, we hypothesized that NMB is related to corticotroph adenoma cell proliferation and hormone secretion. To examine this hypothesis, we investigated the expression of NMB and its receptor NMBR in human corticotroph adenoma and the effects of a NMBR antagonist on AtT-20 cells, a mouse corticotroph adenoma cell line, and patient-derived corticotroph adenoma cells. Methods: 1. NMB and NMBR expression in human pituitary adenoma: We performed real-time qPCR and immunostaining on human pathological specimens of corticotrophs, somatotrophs, and non-functioning pituitary adenoma to investigate NMB and NMBR expression. 2. Experiments in AtT-20 cells: We extracted mRNAs and proteins from AtT-20 cells after incubation with 100nM NMBR antagonist PD168368, and performed real-time qPCR and western blotting analyses to investigate Pomc expression. 3. Experiments in patient-derived corticotroph adenoma cells: We isolated surgically resected human corticotroph adenoma cells from patients who underwent trans-sphenoidal surgery and investigated POMC mRNA expression by real-time qPCR after incubation with PD168368. Statistical analysis: One-way ANOVA was employed to compare values among multiple groups. If the ANOVA revealed significant differences, the Tukey-Kramer post-hoc test was employed to compare values between two specific groups. Dunnett’s post-hoc test was employed to compare values with the control group. Statistical significance was defined as p < 0.05. Results: 1. NMB and NMBR expression levels were significantly higher in human corticotroph adenoma (13 and 33 times higher than non-functioning adenoma, respectively) than in somatotroph adenoma (2 and 3 times higher than non-functioning adenoma, respectively) and non-functioning adenoma in the qPCR analyses. Immunostaining confirmed higher expression of NMB and NMBR in corticotroph adenoma than in somatotroph and non-functioning adenoma. 2. Treatment with 100 nM PD168368 significantly suppressed Pomc mRNA and protein expression in AtT-20 cells by 22%±3% and 25%±10%, respectively. 3. Treatment with 1 µM PD168368 significantly suppressed POMC mRNA expression in human corticotroph adenoma cells by 18%±1%. Conclusions: NMB and NMBR were both expressed in human corticotroph adenoma, suggesting that NMB may stimulate adenoma cell proliferation and hormone secretion in autocrine or paracrine manners. Because the NMBR antagonist suppressed Pomc expression in both AtT-20 cells and human corticotroph adenoma cells, it may represent a potential treatment for Cushing disease. Reference: (1) Kameda H et al., Endocrinology 2014;155(7):2492-9.

Effect of LncRNA BLACAT1 on the Proliferation and Apoptosis of Osteoblasts in Ankylosing Spondylitis Through Regulating RANKL/OPG Signaling Pathway

2019 ◽  
Vol 9 (9) ◽  
pp. 1279-1285
Author(s):  
Hongbin Zhu ◽  
Zongbao Gao ◽  
Tao Wang ◽  
Zhigang Lei ◽  
Maoqin Sun
Keyword(s):  
Cell Proliferation ◽  
Ankylosing Spondylitis ◽  
Disease Activity ◽  
Mrna Expression ◽  
Real Time ◽  
Signaling Pathway ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Stable Phase ◽  
Proliferation And Apoptosis

Ankylosing spondylitis (AS) is an autoimmune disorder. LncRNA BLACAT1 involves in several diseases such as inflammation and immune diseases, but the expression and role of LncRNA BLACAT1 in AS remains unclear. AS patients and controls were selected. LcRNA BLACAT1 expression in peripheral blood was analyzed by Real time PCR, and its correlation with CRP, ESR and AS activity was analyzed. Osteoblast hFOB 1.19 was cultured and transfected with LncRNA BLACAT1 plasmid and si-LncRNA BLACAT1, respectively followed by analysis of cell proliferation by MTT assay, apoptosis by Caspase 3, ADAMTS-4 and Runx2 mRNA expression by Real time PCR, IFN-γ and TNF-α secretion by ELISA, and as RANKL, OPG, and nerve growth factor NGF level by western blot. AS patients presented significantly elevated LcRNA BLACAT1 level than controls (P < 0.05) with higher expression in active AS patients than stable phase (P < 0.05). BLACAT1 was positively associated with CRP and disease activity index (P < 0.05). Overexpression of LncRNA BLACAT1 in osteoblast hFOB 1.19 significantly decreased cell proliferation, elevated Caspase 3 activity, increased ADAMTS-4 mRNA expression, decreased Runx2 mRNA expression, and increased IFN-γ and TNF-α sevretion, and RANKL expression, and decreased OPG and NGF expression (P < 0.05). However, the above changes were reversed by si-LncRNA BLACAT1. LncRNA BLACAT1 expression in AS patients can reflect the degree of disease activity, and its mechanism may be through regulation of RANKL/OPG signaling pathway, which affects the proliferation and apoptosis of osteoblasts in AS.

Role of urocortin 2 secreted by the pituitary in the stress-induced suppression of luteinizing hormone secretion in rats

2010 ◽  
Vol 299 (4) ◽  
pp. E567-E575 ◽  
Author(s):  
Takahiro Nemoto ◽  
Azusa Iwasaki-Sekino ◽  
Naoko Yamauchi ◽  
Tamotsu Shibasaki
Keyword(s):  
Luteinizing Hormone ◽  
Mrna Expression ◽  
Hormone Secretion ◽  
Anterior Lobe ◽  
Intermediate Lobe ◽  
Β Subunit ◽  
Luteinizing Hormone Secretion ◽  
Subunit Mrna ◽  
Pomc Mrna

We have previously shown that urocortin 2 (Ucn 2), a member of the corticotropin-releasing factor (CRF) peptide family that binds to CRF type 2 receptor, is expressed in proopiomelanocortin (POMC) cells of rat pituitary and that its secretion and expression are increased by CRF in both the anterior and intermediate lobes and suppressed by glucocorticoids in the anterior lobe. We have also shown that Ucn 2 secreted by POMC cells acts on gonadotrophs expressing CRF type 2 receptors and inhibits the expression and secretion of gonadotropins. In the present study, we examined whether pituitary Ucn 2 is involved in stress-induced inhibition of gonadotropin secretion. A 90-min period of immobilization stress increased POMC mRNA expression without influencing Ucn 2 mRNA expression and suppressed luteinizing hormone (LH) β-subunit mRNA expression in the anterior lobe and plasma LH levels, while it increased both POMC and Ucn 2 mRNA expression in the intermediate lobe of the pituitary. Pretreatment with anti-CRF IgG blocked immobilization-induced increases in plasma ACTH and corticosterone and in POMC mRNA expression in both pituitary lobes and Ucn 2 mRNA expression in the intermediate pituitary. It also blocked immobilization-induced suppression of plasma LH and LH β-subunit mRNA expression. Pretreatment with anti-Ucn 2 IgG blocked immobilization-induced suppression of plasma LH and LH β-subunit expression without affecting immobilization-induced ACTH and corticosterone release and POMC or Ucn 2 mRNA expression. These results suggest that CRF suppresses the secretion and expression of LH probably through pituitary Ucn 2 in stress.

scholarly journals β-catenin knockdown inhibits pituitary adenoma cell proliferation and invasion via interfering with AKT and gelatinases expression

2015 ◽  
Vol 46 (4) ◽  
pp. 1643-1650 ◽  
Author(s):  
CHENGCHENG ZHAO ◽  
MENG ZHANG ◽  
WENLAN LIU ◽  
CHUANFANG WANG ◽  
QIUSHENG ZHANG ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Pituitary Adenoma ◽  
Adenoma Cell ◽  
Proliferation And Invasion

scholarly journals Effects of Oxytocin on Cell Proliferation in a Corticotroph Adenoma Cell Line

2019 ◽  
Vol 34 (3) ◽  
pp. 302
Author(s):  
Jung Soo Lim ◽  
Young Woo Eom ◽  
Eun Soo Lee ◽  
Hyeong Ju Kwon ◽  
Ja-Young Kwon ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Line ◽  
Corticotroph Adenoma ◽  
Adenoma Cell

Gestational profile of Na+/H+ exchanger and Cl−/HCO3− anion exchanger mRNA expression in placenta using real-time QPCR

Placenta ◽  
2005 ◽  
Vol 26 (1) ◽  
pp. 93-98 ◽  
Author(s):  
H.A. Lacey ◽  
T. Nolan ◽  
S.L. Greenwood ◽  
J.D. Glazier ◽  
C.P. Sibley
Keyword(s):  
Mrna Expression ◽  
Real Time ◽  
Anion Exchanger ◽  
Real Time Qpcr

PI3K/Akt/mTOR pathway involvement in regulating growth hormone secretion in a rat pituitary adenoma cell line

Endocrine ◽  
2017 ◽  
Vol 60 (2) ◽  
pp. 308-316 ◽  
Author(s):  
Carmelina Di Pasquale ◽  
Erica Gentilin ◽  
Simona Falletta ◽  
Mariaenrica Bellio ◽  
Mattia Buratto ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Pituitary Adenoma ◽  
Cell Line ◽  
Hormone Secretion ◽  
Growth Hormone Secretion ◽  
Mtor Pathway ◽  
Adenoma Cell ◽  
Rat Pituitary

Effect of cortisol on neurophysin I/oxytocin and peptidyl glycine-α-amidating mono-oxygenase mRNA expression in bovine luteal and granulosa cells

2013 ◽  
Vol 16 (2) ◽  
pp. 231-239
Author(s):  
A. Ziolkowska ◽  
J. Mlynarczuk ◽  
J. Kotwica
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Molecular Mechanisms ◽  
Hormone Secretion ◽  
Oestrous Cycle ◽  
Luteal Cells ◽  
Ru 486 ◽  
Ovarian Cells ◽  
Key Genes ◽  
Ovary Function

Abstract Cortisol stimulates the synthesis and secretion of oxytocin (OT) from bovine granulosa and luteal cells, but the molecular mechanisms of cortisol action remain unknown. In this study, granulosa cells or luteal cells from days 1-5 and 11-15 of the oestrous cycle were incubated for 4 or 8 h with cortisol (1x10-5, 1x10-7 M). After testing cell viability and hormone secretion (OT, progesterone, estradiol), we studied the effect of cortisol on mRNA expression for precursor of OT (NP-I/OT) and peptidyl glycine-α-amidating mono-oxygenase (PGA). The influence of RU 486 (1x10-5 M), a progesterone receptor blocker and inhibitor of the glucocorticosteroid receptor (GR), on the expression for both genes was tested. Cortisol increased the mRNA expression for NP-I/OT and PGA in granulosa cells and stimulated the expression for NP-I/OT mRNA in luteal cells obtained from days 1-5 and days 11-15 of the oestrous cycle. Expression for PGA mRNA was increased only in luteal cells from days 11-15 of the oestrous cycle. In addition, RU 486 blocked the cortisol-stimulated mRNA expression for NP-I/OT and PGA in both types of cells. These data suggest that cortisol affects OT synthesis and secretion in bovine ovarian cells, by acting on the expression of key genes, that may impair ovary function.

scholarly journals Follicle-Stimulating Hormone Inhibits Adenosine 5′-Monophosphate-Activated Protein Kinase Activation and Promotes Cell Proliferation of Primary Granulosa Cells in Culture through an Akt-Dependent Pathway

Endocrinology ◽  
2009 ◽  
Vol 150 (2) ◽  
pp. 929-935 ◽  
Author(s):  
Pradeep P. Kayampilly ◽  
K. M. J. Menon
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Protein Kinase ◽  
Mrna Expression ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Ampk Activation ◽  
Cyclin D2 ◽  
Cell Cycle Inhibitor ◽  
Dependent Pathway

FSH, acting through multiple signaling pathways, regulates the proliferation and growth of granulosa cells, which are critical for ovulation. The present study investigated whether AMP-activated protein kinase (AMPK), which controls the energy balance of the cell, plays a role in FSH-mediated increase in granulosa cell proliferation. Cells isolated from immature rat ovaries were grown in serum-free, phenol red free DMEM-F12 and were treated with FSH (50 ng/ml) for 0, 5, and 15 min. Western blot analysis showed a significant reduction in AMPK activation as observed by a reduction of phosphorylation at thr 172 in response to FSH treatment at all time points tested. FSH also reduced AMPK phosphorylation in a dose-dependent manner with maximum inhibition at 100 ng/ml. The chemical activator of AMPK (5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside, 0.5 mm) increased the cell cycle inhibitor p27 kip expression significantly, whereas the AMPK inhibitor (compound C, 20 μm) and FSH reduced p27kip expression significantly compared with control. FSH treatment resulted in an increase in the phosphorylation of AMPK at ser 485/491 and a reduction in thr 172 phosphorylation. Inhibition of Akt phosphorylation using Akt inhibitor VIII reversed the inhibitory effect of FSH on thr 172 phosphorylation of AMPK, whereas ERK inhibitor U0126 had no effect. These results show that FSH, through an Akt-dependent pathway, phosphorylates AMPK at ser 481/495 and inhibits its activation by reducing thr 172 phosphorylation. AMPK activation by 5-amino-imidazole-4-carboxamide-1-β-d-ribofuranoside treatment resulted in a reduction of cell cycle regulatory protein cyclin D2 mRNA expression, whereas FSH increased the expression by 2-fold. These results suggest that FSH promotes granulosa cell proliferation by increasing cyclin D2 mRNA expression and by reducing p27 kip expression by inhibiting AMPK activation through an Akt-dependent pathway. FSH stimulates granulosa cell proliferation by reducing cell cycle inhibitor p27 kip through AMP kinase inhibition.

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