The cAMP pathway promotes sirtuin-1 expression in human granulosa-lutein cells

Szymańska, Magdalena; Manthe, Sarah; Shrestha, Ketan; Girsh, Eliezer; Harlev, Avi; Meidan, Rina

doi:10.1016/j.repbio.2020.07.010

Cited by 8 publications

(38 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Total proteins were extracted from cultured cells as described previously [12,46]. Briefly, cells were isolated by scrapping cells in sample buffer (1 mol/L Tris-HCl (pH 6.8), 20% (v/v) glycerol, 4% (w/v) SDS and 0.0002% (w/v) bromophenol blue and 1% (v/v) β-mercaptoethanol) containing phosphatase inhibitor cocktail 1 (Sigma Aldrich, St. Louis, MO, USA).…”

Section: Protein Extraction and Western Blot Analysesmentioning

confidence: 99%

“…Along with ovulatory genes, LH induces two key luteal angiogenic factors: fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor A (VEGFA) [9,10]; expressed at particularly high levels during follicular-luteal transition. An additional factor highly expressed in a cAMPdependent manner in luteinized human GCs (hGLCs) is sirtuin-1 (SIRT1) [11][12][13]. SIRT1 is a NAD + -dependent deacetylase associated with various biological functions [14][15][16], including regulation of proliferation and secretory activity of GCs [17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

“…SIRT1 is a NAD + -dependent deacetylase associated with various biological functions [14][15][16], including regulation of proliferation and secretory activity of GCs [17][18][19][20]. It was further demonstrated that SIRT1 elevated cAMP on its own and additively with human chorionic gonadotropin (hCG) [12], however whether SIRT1 affect ovulatory and angiogenic genes is yet to be examined.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Diverse actions of sirtuin-1 on ovulatory genes and cell death pathways in human granulosa cells

Sapuleni

Szymańska

Meidan

2022

Reprod Biol Endocrinol

Self Cite

View full text Add to dashboard Cite

Background Human granulosa-lutein cells (hGLCs) amply express sirtuin-1 (SIRT1), a NAD + -dependent deacetylase that is associated with various cellular functions. SIRT1 was shown to elevate cAMP on its own and additively with human chorionic gonadotropin (hCG), it is therefore interesting to examine if SIRT1 affects other essential hGLC functions. Methods Primary hGLCs, obtained from the follicular aspirates of women undergoing IVF and SV40-transfected, immortalized hGLCs (SVOG cells), were used. Primary cells were treated with SIRT1 specific activator SRT2104, as well as hCG or their combination. Additionally, siRNA-targeting SIRT1 construct was used to silence endogenous SIRT1 in SVOG cells. PTGS2, EREG, VEGFA and FGF2 expression was determined using quantitative polymerase chain reaction (qPCR). Apoptotic and necroptotic proteins were determined by specific antibodies in western blotting. Cell viability/apoptosis was determined by the XTT and flow cytometry analyses. Data were analyzed using student t-test or Mann–Whitney U test or one-way ANOVA followed by Tukey HSD post hoc test. Results In primary and immortalized hGLCs, SRT2104 significantly upregulated key ovulatory and angiogenic genes: PTGS2, EREG, FGF2 and VEGFA, these effects tended to be further augmented in the presence of hCG. Additionally, SRT2104 dose and time-dependently decreased viable cell numbers. Flow cytometry of Annexin V stained cells confirmed that SIRT1 reduced live cell numbers and increased late apoptotic and necrotic cells. Moreover, we found that SIRT1 markedly reduced anti-apoptotic BCL-XL and MCL1 protein levels and increased cleaved forms of pro-apoptotic proteins caspase-3 and PARP. SIRT1 also significantly induced necroptotic proteins RIPK1 and MLKL. RIPK1 inhibitor, necrostatin-1 mitigated SIRT1 actions on RIPK1 and MLKL but also on cleaved caspase-3 and PARP and in accordance on live and apoptotic cells, implying a role for RIPK1 in SIRT1-induced cell death. SIRT1 silencing produced inverse effects on sorted cell populations, anti-apoptotic, pro-apoptotic and necroptotic proteins, corroborating SIRT1 activation. Conclusions These findings reveal that in hGLCs, SIRT1 enhances the expression of ovulatory and angiogenic genes while eventually advancing cell death pathways. Interestingly, these seemingly contradictory events may have occurred in a cAMP-dependent manner.

show abstract

Section: Protein Extraction and Western Blot Analysesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Diverse actions of sirtuin-1 on ovulatory genes and cell death pathways in human granulosa cells

Sapuleni

Szymańska

Meidan

2022

Reprod Biol Endocrinol

Self Cite

View full text Add to dashboard Cite

show abstract

“…The transfection was performed according to the manufacturer's instructions using Opti-MEM (31985070, ThermoFisher Scientific) for complex dilution. The cells were transfected with 10 nM siRNA for the SIRT1 (sequence, [10]) and SIRT3 (Santa Cruz) targets. For every transfection, a nontargeting (NT) siRNA was used as a control (Silencer Select Negative Control #1, ThermoFisher Scientific).…”

Section: Sirna Transfectionmentioning

confidence: 99%

“…Evidence has accumulated specifically pinpointing SIRT1 in the control of GC proliferation, apoptosis and secretory activity [3,4,8,9]. Recently, a role in luteinized human GCs was addressed, and for example, evidence for a cAMP-driven expression of SIRT1 was provided [10,11]. Additionally, the SIRT-expression profiles in these cells differ between normal donors and infertile women [12], suggesting involvement in functions related to fertility and infertility.…”

Section: Introductionmentioning

confidence: 99%

Sirtuin 1 and Sirtuin 3 in Granulosa Cell Tumors

Schmid¹,

Dietrich²,

Forné³

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

Sirtuins (SIRTs) are NAD+-dependent deacetylases that regulate proliferation and cell death. In the human ovary, granulosa cells express sirtuin 1 (SIRT1), which has also been detected in human tumors derived from granulosa cells, i.e., granulosa cell tumors (GCTs), and in KGN cells. KGN cells are an established cellular model for the majority of GCTs and were used to explore the role of SIRT1. The SIRT1 activator SRT2104 increased cell proliferation. By contrast, the inhibitor EX527 reduced cell numbers, without inducing apoptosis. These results were supported by the outcome of siRNA-mediated silencing studies. A tissue microarray containing 92 GCTs revealed nuclear and/or cytoplasmic SIRT1 staining in the majority of the samples, and also, SIRT2-7 were detected in most samples. The expression of SIRT1–7 was not correlated with the survival of the patients; however, SIRT3 and SIRT7 expression was significantly correlated with the proliferation marker Ki-67, implying roles in tumor cell proliferation. SIRT3 was identified by a proteomic analysis as the most abundant SIRT in KGN. The results of the siRNA-silencing experiments indicate involvement of SIRT3 in proliferation. Thus, several SIRTs are expressed by GCTs, and SIRT1 and SIRT3 are involved in the growth regulation of KGN. If transferable to GCTs in situ, these SIRTs may represent novel drug targets.

show abstract

Emerging roles of SIRT1 activator, SRT2104, in disease treatment

Chang,

Li,

Lin

et al. 2024

Sci Rep

View full text Add to dashboard Cite

Silent information regulator 1 (SIRT1) is a NAD+-dependent class III deacetylase that plays important roles in the pathogenesis of numerous diseases, positioning it as a prime candidate for therapeutic intervention. Among its modulators, SRT2104 emerges as the most specific small molecule activator of SIRT1, currently advancing into the clinical translation phase. The primary objective of this review is to evaluate the emerging roles of SRT2104, and to explore its potential as a therapeutic agent in various diseases. In the present review, we systematically summarized the findings from an extensive array of literature sources including the progress of its application in disease treatment and its potential molecular mechanisms by reviewing the literature published in databases such as PubMed, Web of Science, and the World Health Organization International Clinical Trials Registry Platform. We focuses on the strides made in employing SRT2104 for disease treatment, elucidating its potential molecular underpinnings based on preclinical and clinical research data. The findings reveal that SRT2104, as a potent SIRT1 activator, holds considerable therapeutic potential, particularly in modulating metabolic and longevity-related pathways. This review establishes SRT2104 as a leading SIRT1 activator with significant therapeutic promise.

show abstract

The cAMP pathway promotes sirtuin-1 expression in human granulosa-lutein cells

Cited by 8 publications

References 60 publications

Diverse actions of sirtuin-1 on ovulatory genes and cell death pathways in human granulosa cells

Diverse actions of sirtuin-1 on ovulatory genes and cell death pathways in human granulosa cells

Sirtuin 1 and Sirtuin 3 in Granulosa Cell Tumors

Emerging roles of SIRT1 activator, SRT2104, in disease treatment

Contact Info

Product

Resources

About