Elevated GnRH receptor expression plus GnRH agonist treatment inhibits the growth of a subset of papillomavirus 18‐immortalized human prostate cells

Morgan, Kevin; Stavrou, Emmanouil; Leighton, Samuel P; Miller, Nicola; Sellar, Robin; Millar, Robert P.

doi:10.1002/pros.21308

Cited by 13 publications

(20 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MCF-7, MDA-MB-231, ZR-75-1, and T47D cells were from American Type Culture Collection (LGC, UK). The GnRH receptor stably transfected HEK293 [SCL60] and prostate WPE-1-NB26-8 cell lines described elsewhere [8,9] together with HEK293 cells were used as controls for comparison. These transfected models have previously been shown to demonstrate growth responses to triptorelin [8,9].…”

Section: Methodsmentioning

confidence: 99%

“…Medium for SVCT cells was supplemented with recombinant human insulin and hydrocortisone as specified by the suppliers (ECACC, UK). HEK293 [SCL60] and WPE-1-NB26-8 cells were cultured as described elsewhere [9]. Cells were transfected with a plasmid construct, pcDNA3.1(+) (neo) (Invitrogen, UK) containing a rat GnRH receptor cDNA insert, using Fugene 6 (Roche, UK) in Optimem-I (GIBCO, Invitrogen, UK).…”

Section: Methodsmentioning

confidence: 99%

“…Levels of GnRH receptor at the cell surface were measured as described elsewhere, using 125 I-labeled His 5 D-Tyr 6 GnRH-I as a radiotracer [8,9]. Cells were grown in 12 or 24 well plastic culture plates.…”

Section: Methodsmentioning

confidence: 99%

“…Cells were seeded into 12 well plates and growth was monitored using the sulforhodamine B (SRB) staining assay described previously [8,9]. Two milliliters culture medium per well was sufficient to sustain cell growth over all time courses investigated.…”

Section: Methodsmentioning

confidence: 99%

“…Cell-type specific features influencing GnRH receptor signaling and cell growth-inhibition have been described in cell lines stably expressing elevated levels of the GnRH receptor [8-10]. So far, the ability of GnRH agonist to inhibit cell growth appears to correlate with the level of GnRH receptor expression at the cell surface and with the magnitude of inositol phosphate production elicited by receptor activation [8,9]. GnRH receptor activation coupled to Gα q/11 -Gβγ proteins leads to elevation of intracellular Ca 2+ levels, altered cytoskeletal function and changes in protein kinase activity, including protein kinase C (PKC), mitogen activated serine/threonine kinases (MAPkinases, MAPK) and stress-activated kinases Cell-type specific effects of GnRH receptor activation on levels of phosphorylated-ERK1/2 (p-ERK1/2) have been observed [8,9] which probably reflect the complexity of protein scaffolds interacting with and influencing MAPK.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

GnRH receptor activation competes at a low level with growth signaling in stably transfected human breast cell lines

et al. 2011

Self Cite

View full text Add to dashboard Cite

BackgroundGonadotrophin releasing hormone (GnRH) analogs lower estrogen levels in pre-menopausal breast cancer patients. GnRH receptor (GnRH-R) activation also directly inhibits the growth of certain cells. The applicability of GnRH anti-proliferation to breast cancer was therefore analyzed.MethodsGnRH-R expression in 298 primary breast cancer samples was measured by quantitative immunofluorescence. Levels of functional GnRH-R in breast-derived cell lines were assessed using 125I-ligand binding and stimulation of 3H-inositol phosphate production. Elevated levels of GnRH-R were stably expressed in cells by transfection. Effects of receptor activation on in vitro cell growth were investigated in comparison with IGF-I and EGF receptor inhibition, and correlated with intracellular signaling using western blotting.ResultsGnRH-R immunoscoring was highest in hormone receptor (triple) negative and grade 3 breast tumors. However prior to transfection, functional endogenous GnRH-R were undetectable in four commonly studied breast cancer cell lines (MCF-7, ZR-75-1, T47D and MDA-MB-231). After transfection with GnRH-R, high levels of cell surface GnRH-R were detected in SVCT and MDA-MB-231 clones while low-moderate levels of GnRH-R occurred in MCF-7 clones and ZR-75-1 clones. MCF-7 sub-clones with high levels of GnRH-R were isolated following hygromycin phosphotransferase transfection. High level cell surface GnRH-R enabled induction of high levels of 3H-inositol phosphate and modest growth-inhibition in SVCT cells. In contrast, growth of MCF-7, ZR-75-1 or MDA-MB-231 clones was unaffected by GnRH-R activation. Cell growth was inhibited by IGF-I or EGF receptor inhibitors. IGF-I receptor inhibitor lowered levels of p-ERK1/2 in MCF-7 clones. Washout of IGF-I receptor inhibitor resulted in transient hyper-elevation of p-ERK1/2, but co-addition of GnRH-R agonist did not alter the dynamics of ERK1/2 re-phosphorylation.ConclusionsBreast cancers exhibit a range of GnRH-R immunostaining, with higher levels of expression found in triple-negative and grade 3 cancers. However, functional cell surface receptors are rare in cultured cells. Intense GnRH-R signaling in transfected breast cancer cells did not markedly inhibit growth, in contrast to transfected HEK 293 cells indicating the importance of intracellular context. GnRH-R signaling could not counteract IGF-I receptor-tyrosine kinase addiction in MCF-7 cells. These results suggest that combinatorial strategies with growth factor inhibitors will be needed to enhance GnRH anti-proliferative effects in breast cancer

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

GnRH receptor activation competes at a low level with growth signaling in stably transfected human breast cell lines

et al. 2011

Self Cite

View full text Add to dashboard Cite

show abstract

Gonadotropin-releasing hormone receptors as molecular therapeutic targets in prostate cancer: Current options and emerging strategies

Limonta

Manea

2013

Cancer Treatment Reviews

View full text Add to dashboard Cite

Gonadotropin-releasing hormone receptor inhibits triple-negative breast cancer proliferation and metastasis

Chen¹,

2022

J Int Med Res

View full text Add to dashboard Cite

Background Gonadotropin-releasing hormone receptor (GnRHR) is expressed in several malignant tumors and inhibits the proliferation and metastasis of cancer cells, but its role in triple-negative breast cancers (TNBCs) is unclear. This study investigated the biological effects of GnRHR and their influence on TNBC prognosis. Methods The GSE21653 database was used to obtain information about GnRHR expression and clinicopathological factors in patients with TNBC. GnRHR was activated in cultured MDA-MB-231 and MDA-MB-468 cells by leuprolide acetate and antagonized by elagolix sodium. Cell proliferation was assessed by the cell counting kit-8 and colony formation assays. Cell metastasis was detected by the wound healing assay and Transwell assay. Apoptosis and the cell cycle were investigated by flow cytometry. GnRHR protein expression was determined by western blotting. Results GnRHR mRNA expression was significantly higher in patients with TNBC than in hormone receptor+/human epidermal growth factor receptor (HER)2– and HER2+ patients with breast cancer. Patients with high GnRHR expression had significantly better disease-free survival than those with lower expression. Activated GnRHR significantly inhibited cell proliferation and metastasis, increased apoptosis, and enhanced GnRHR protein expression levels. Conclusion GnRHR inhibits TNBC proliferation and metastasis, suggesting it could be targeted for TNBC treatment.

show abstract

Elevated GnRH receptor expression plus GnRH agonist treatment inhibits the growth of a subset of papillomavirus 18‐immortalized human prostate cells

Cited by 13 publications

References 63 publications

GnRH receptor activation competes at a low level with growth signaling in stably transfected human breast cell lines

GnRH receptor activation competes at a low level with growth signaling in stably transfected human breast cell lines

Gonadotropin-releasing hormone receptors as molecular therapeutic targets in prostate cancer: Current options and emerging strategies

Gonadotropin-releasing hormone receptor inhibits triple-negative breast cancer proliferation and metastasis

Contact Info

Product

Resources

About