GH3 and RC-4BC cell lines are not suitable as in vitro models to study prolactin modulation and AHR responsiveness in rat pituitary

Brand, A.; Rubinstein, Efrat; Berg, Martin van den; Duursen, Majorie B.M. van

doi:10.1016/j.mce.2019.110520

Cited by 10 publications

(4 citation statements)

References 40 publications

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“…For testing the effects of NESF or NLP on GH, cells at 90% of confluence were used. The growth culture media was supplemented with rat NESF (custom synthesized peptide 6 , > 95% purity, ABGENT, USA) or rat NLP (custom synthesized peptide 7 , > 95% purity, ABGENT) at 0 (Control), 0.001, 0.01, 0.1, 1, 10, 100 or 1000 nM concentration and cells were incubated for 1, 6 or 24 h. The 24 h time point was chosen as previous studies with the same cell lines reported mRNA changes at that time 38 , 39 . After incubation, cells were washed twice with 1X PBS before the collection of total RNA or the protein content, as explained below.…”

Section: Methodsmentioning

confidence: 99%

Nesfatin-1 and nesfatin-1-like peptide suppress growth hormone synthesis via the AC/PKA/CREB pathway in mammalian somatotrophs

Vélez

Unniappan

2020

Sci Rep

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Nesfatin-1 (NESF) and NESF-like peptide (NLP), encoded in nucleobindin 2 and 1 (NUCB2 and NUCB1), respectively, are orphan ligands and metabolic factors. We hypothesized that NESF and NLP suppress growth hormone (GH) synthesis, and aimed to determine whether mammalian somatotrophs are a source and site of action of these peptides. Using immortalized rat somatotrophs (GH3 cells), NUCB expression was determined by qPCR, immunofluorescence and Western blot. NESF and NLP binding to GH3 cells was tested using fluorescence imaging. Both time- and concentration-dependent studies were performed to test whether NESF and NLP affect GH. Moreover, the ability of these peptides to modulate the effects of ghrelin, and cell-signaling pathways were studied. GH3 cells express NUCB mRNAs and protein. Labeled NESF and NLP bind to the surface of GH3 cells, and incubation with either NESF or NLP decreased GH mRNA and protein expression, downregulated pit-1 mRNA, and blocked the GH stimulatory effects of ghrelin. Pre-incubation with either of these peptides reduced CREB phosphorylation by an AC-activator, but not when PKA was directly activated by a cAMP analog. Our results indicate that rat somatotrophs are a source of NUCBs, and that NESF and NLP downregulate GH synthesis through the AC/PKA/CREB signaling pathway.

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Section: Methodsmentioning

confidence: 99%

Nesfatin-1 and nesfatin-1-like peptide suppress growth hormone synthesis via the AC/PKA/CREB pathway in mammalian somatotrophs

Vélez

Unniappan

2020

Sci Rep

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“…It contains all types of anterior pituitary cells with majority of cells containing FSH, LH, and prolactin, with many bihormonal cells within cell line [15]. The cell line was used to study basal prolactin secretion in comparison with primary pituitary cells and other aspects of pituitary cell functionality [16,17]. They are suitable to address questions related to pituitary adenoma growth.…”

Section: Discussionmentioning

confidence: 99%

RC-4BC cells express nicotinic and muscarinic acetylcholine receptors

Zhubanova

Filchakova

2022

PLoS ONE

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Acetylcholine is one of the most important endogenous neurotransmitters in a range of organisms spanning different animal phyla. Within pituitary gland it acts as autocrine and paracrine signal. In a current study we assessed expression profile of the different subunits of nicotinic as well as muscarinic acetylcholine receptors in RC-4BC cells, which are derived from rat pituitary gland tumor. Our findings indicate that β2, δ, and M2 subunits are expressed by the cells with the lowest Ct values compared to other tested subunits. The detected Ct values were 26.6±0.16, 27.95±0.5, and 28.8±0.25 for β2, δ, and M2 subunits, respectively.

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“…Isolation of primary pituitary cells were performed as previously described [43]. In brief, human pituitary adenoma and rat normal pituitary tissues were minced and incubated with 0.5% collagenase type I and 0.05% DNase I (Solarbio, Beijing, China) at 37℃ for 45 min.…”

Section: Isolation and Culture Of Primary Pituitary Cellsmentioning

confidence: 99%

PER2 integrates circadian disruption and pituitary tumorigenesis

Guo¹,

Cen²,

Weng³

et al. 2023

Theranostics

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Rationale:The role of circadian clock in pituitary tumorigenesis remains elusive. Here we investigate whether and how circadian clock modulates the development of pituitary adenomas. Methods and Results: We found altered expression of pituitary clock genes in patients with pituitary adenomas. In particular, PER2 is prominently upregulated. Further, jetlagged mice with PER2 upregulation have accelerated growth of GH3 xenograft tumor. Conversely, loss of Per2 protects mice against developing estrogen-induced pituitary adenoma. Similar antitumor effect is observed for SR8278, a chemical that can decrease pituitary PER2 expression. RNA-seq analysis suggests involvement of cell cycle disturbance in PER2 regulation of pituitary adenoma. Subsequent in vivo and cell-based experiments validate that PER2 induces pituitary expression of Ccnb2, Cdc20 and Espl1 (three cell cycle genes) to facilitate cell cycle progression and inhibit apoptosis, thereby promoting pituitary tumorigenesis. Mechanistically, PER2 regulates the transcription of Ccnb2, Cdc20 and Espl1 through enhancing the transcriptional activity of HIF-1α. HIF-1α trans-activates Ccnb2, Cdc20 and Espl1 via direct binding to its specific response element in the gene promoters. Conclusion: PER2 integrates circadian disruption and pituitary tumorigenesis. These findings advance our understanding of crosstalk between circadian clock and pituitary adenomas and highlight the relevance of clock-based approaches in disease management.

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GH3 and RC-4BC cell lines are not suitable as in vitro models to study prolactin modulation and AHR responsiveness in rat pituitary

Cited by 10 publications

References 40 publications

Nesfatin-1 and nesfatin-1-like peptide suppress growth hormone synthesis via the AC/PKA/CREB pathway in mammalian somatotrophs

Nesfatin-1 and nesfatin-1-like peptide suppress growth hormone synthesis via the AC/PKA/CREB pathway in mammalian somatotrophs

RC-4BC cells express nicotinic and muscarinic acetylcholine receptors

PER2 integrates circadian disruption and pituitary tumorigenesis

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