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Proteome alterations in the human granulosa-like tumor cell line KGN after CRISPR/Cas9-based deletion of TRPV2

ABSTRACT: Human ovarian granulosa cell tumors (GCT) express the cation channel transient receptor potential vanilloid 2 (TRPV2) to a large percentage (95%), as detected by analyzing a panel of 72 tissue samples. To examine its role, we turned to the GCT-derived cell line KGN. CRISPR/Cas9-based deletion of TRPV2 resulted in alterations in the proteome of these cells.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Thomas Fröhlich

LAB HEAD: Thomas Fröhlich

PROVIDER: PXD060778 | Pride | 2026-02-17

REPOSITORIES: Pride

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			Action	DRS
	Clone4_TRPV2_1.raw	Raw
	Clone4_TRPV2_2.raw	Raw
	Clone4_TRPV2_3.raw	Raw
	Clone4_TRPV2_4.raw	Raw
	Clone4_TRPV2_5.raw	Raw

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Publications

TRPV2 regulates cell fate in the human granulosa-like tumor cell line KGN: implications for granulosa cell tumors and cannabidiol.

Eubler Katja K Jeevanandan Sree Priyanka SP Caban Karolina Magdalena KM Stöckl Jan Bernd JB Braun Malte Benjamin MB Herrmann Carola C Schneider Michaela M Kreitmair Nicole N Scholz Lina L Mayr Doris D Renkawitz Jörg J Welter Harald H Müller-Taubenberger Annette A Fröhlich Thomas T Mayerhofer Artur A

Cell communication and signaling : CCS 20260212 1

BACKGROUND: The transient receptor potential vanilloid 2 (TRPV2) channel is known to have strong species-dependent activation modes and divers functions. Based on previous results, which showed expression by human ovarian granulosa cells, we studied human TRPV2 in human granulosa cell tumors (GCTs) and derived cells (KGN cells). GCTs are rare ovarian tumors, for which neither specific therapies, nor adequate markers are available. METHODS: We analyzed primary GCTs, including a panel of 63 GCT sa ...[more]

PMID: 41680789

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Proteome alterations in Filamin A deficient granulosa cell tumor-derived KGN cells.

Project description:The goal of the study was to evaluate consequences of Filamin A deficiency in granulosa cell tumor-derived KGN cells. Therefore, proteomes of WT and Filamin A deficient KGN cells were quantitatively compared using LC-MS/MS.

2026-05-22 | PXD064136 | Pride

Role of YAP/TAZ in KGN human granulosa cell line

Project description:To identify the YAP/TAZ target genes in granulosa cells, DNA microarray experiments were performed in KGN cells where YAP/TAZ were knocked down. The effect of the presence or absence of 8Br-cAMP, an intracellular second messenger of FSH, was also investigated in KGN cells.

2022-11-04 | GSE203080 | GEO

Role of YAP/TAZ in KGN human granulosa cell line [Clariom_S_Human]

2022-11-04 | GSE213916 | GEO

Mitochondrial dynamics in ovarian granulosa cells during follicular growth and in human KGN cells

Project description:In the ovary, proliferation and differentiation of granulosa cells (GCs) drive the growth of follicles. This is, in part, dependent on gonadotropins. Our immunohistochemical studies provided hints of mitochondrial biogenesis and intracellular redistribution in GCs of growing follicles. A cellular model, human KGN cells (granulosa cell tumor cells, derived from growing follicles) was used to study aspects of mitochondrial dynamics. To elevate cAMP and thereby mimic gonadotropin actions, forskolin (FSK) was used, which increased KGN cell size and mitochondrial DNA within 24 h. MitoTracker experiments and ultrastructural 3D reconstruction revealed that FSK treatment induced the formation of elaborate mitochondrial networks. H89, a protein kinase A (PKA) inhibitor, reduced network formation. A proteomic analysis indicated that FSK among others elevated levels of regulators of the cytoskeleton and the steroidogenic enzyme CYP11A1, located in mitochondria, was more than 3-fold increased, implying that cAMP/PKA-associated structural changes go in parallel with the acquisition of steroidogenic competence of mitochondria in KGN cells. In summary, in situ observations showed increases in mitochondria and suggested intracellular trafficking in GCs during follicular growth and indicated that they may partially be under the control of gonadotropins/cAMP. In line with this, elevation of cAMP in KGN profoundly affected mitochondria dynamics in a PKA-dependent manner and implicated cytoskeletal changes.

2022-10-14 | PXD032160 | Pride

Gene knockdown and overexpression of 402C>G FOXL2 in COV434 and KGN cells

Project description:Despite their distinct biology, granulosa cell tumours (GCTs) are treated the same as other ovarian tumours. Intriguingly, a recurring somatic mutation in the transcription factor Forkhead Box L2 (FOXL2) 402C>G has been found in nearly all GCTs examined. This investigation aims to identify the pathogenicity of mutant FOXL2 by studying its altered transcriptional targets. The expression of mutant FOXL2 was reduced in the GCT cell line KGN, and wildtype and mutant FOXL2 were overexpressed in the GCT cell line COV434. Comparisons were made between the transcriptomes of control cells and cells altered by FOXL2 knockdown and overexpression, to detect potential transcriptional targets of mutant FOXL2. Comparisons were made between the transcriptomes of control cells and cells altered by FOXL2 knockdown and overexpression, to detect potential transcriptional targets of mutant FOXL2. Mutant FOXL2 was silenced in KGN cells with the use of siRNA and a non-targetting siRNA control. Wildtype and mutant FOXL2 were overexpressed in COV434 cells with the use of overexpression plasmids and an empty plasmid control. Each experiment was performed in triplicate. Total RNA was harvested at 24h post transfection and hybridised to Affymetrix U133 plus 2.0 arrays.

2012-11-05 | E-GEOD-39890 | biostudies-arrayexpress

Gene knockdown and overexpression of 402C>G FOXL2 in COV434 and KGN cells

2012-11-05 | GSE39890 | GEO

ChIP-chip by array of human adult ovarian granulosa cell tumor derived cell line KGN using anti-FOXL2 antibodies

Project description:ChIP-on-Chip experiment using chromatin from the human adult ovarian granulosa cell tumor derived cell line KGN (Nishi et al, 2001), or from a KGN subclone overexpressing WT FOXL2 (KF3 subclone), and an isovolumic blend of our custom anti-FOXL2 polyclonal antibodies (Cocquet J et al, 2002). Non precipitated sheared matched deproteinized chromatin (Input) was used a control to estimate enrichment peaks (and thus FOXL2 binding sites) from IPed DNA. DNA from three independent ChIP assays (Input extractions) was pooled, and 100ng of DNA was linearly amplified using the Whole Genome Amplification kit (Sigma). The two ChIP-chip experiments serve as biological replicates. Moreover, 42 out of 48 targets promoters chosen from the âenrichment peakâ list were subsequently confirmed as enrichment sites in anti-FOXL2 ChIPed DNA for subsequent experiments.

2011-02-22 | E-MTAB-399 | biostudies-arrayexpress

Melatonin signaling pathways implicated in metabolic processes in human granulosa cells (KGN)

Project description:Female reproduction depends on the metabolic status especially during the period of folliculogenesis. Even though it is believed that melatonin can improve oocyte competence, there is still limited knowledge of how it can modulate metabolic processes during folliculogenesis, and of which signaling pathways are involved in regulating gene expression. To investigate the effects of melatonin on metabolic signals during the antral stage of follicular development, human granulosa-like tumour cells (KGN) were treated with melatonin or forskolin and gene expression was analyzed with RNA seq technology. Following appropriate normalization and the application of a fold change cut-off of 1.5 (FC 1.5, P ≤ 0.05), 1,009 and 922 genes were identified as differentially expressed in response to melatonin and forskolin, respectively. Analysis of major upstream regulators suggested that melatonin may activate PKB/mTOR signaling pathways to program the metabolism of KGN cells to support slower growth and differentiation and to prevent follicular atresia. Similarly, PKA activation through stimulation of cAMP synthesis with FSK seemed to exert the same effects as melatonin in reducing follicular growth and regulating differentiation. This study suggests that it is possible that melatonin acts through PKA and PKB simultaneously in human granulosa cells to prevent follicular atresia and early luteinisation at the antral stage.

2022-03-09 | GSE189124 | GEO

FOXO1/3 and PTEN Depletion in Granulosa Cells Promotes Ovarian Granulosa Cell Tumor Development

Project description:The Forkhead Box, FOXO1 and FOXO3, transcription factors regulate multiple functions in mammalian cells. Selective inactivation of the Foxo1 and Foxo3 genes in murine ovarian granulosa cells severely impairs follicular development and apoptosis causing infertility, and as shown herein, granulosa cell tumor (GCT) formation. Coordinate depletion of the tumor suppressor Pten gene in the Foxo1/3 strain enhanced the penetrance and onset of GCT formation A direct comparison of ovarian granulosa cells from wild type d25 and FOXO/PTEN knockout granulosa cell tumors.

2015-06-17 | E-GEOD-67662 | biostudies-arrayexpress

Histone deacetylase inhibitor, panobinostat, exerts anti-proliferative effect with partial normalization from aberrant epigenetic states on granulosa cell tumor cell lines

Project description:The prognosis of the patients with inoperable or advanced granulosa cell tumors (GCTs) is still poor, and therefore it is important to establish a novel treatment strategy. Here we investigated the in vitro effects of a histone deacetylase inhibitor, panobinostat (PS) on two GCT cell lines (KGN and COV434). GCT cell lines were found to be susceptible to PS treatment and it inhibited cell growth mainly by apoptosis. In cell cycle analysis, PS reduced only the ratio of S phase in GCT cell lines. Combined treatment of PS with a deubiquitinase inhibitor, VLX1570 enhanced the expression of p21, cleaved PARP, cleaved caspase-9, heme oxygenase-1, and the acetylation of histone H4 and α-tubulin, leading to an additive anti-proliferative effect on KGN and COV434. The gene set enrichment analysis revealed that PS treatment suppressed DNA replication- or cell cycle-related gene expression which led to chemotherapeutic cell death and in addition, this treatment induced activation of the gene set of adherens junction towards a normalized direction as well as activation of neuron-related gene sets that might imply unexpected differentiation potential due to epigenetic modification by a HDAC inhibitor in KGN cells. Exposure of KGN and COV434 cells to PS increased the expression of E-cadherin, one of the principal regulators associated with adherens junction in quantitative RT-PCR and immunoblotting analysis. In the present study, we indicate a basis of a novel therapeutic availability of a HDAC inhibitor for the treatment of GCTs and further investigations will be warranted.

2022-04-22 | GSE201203 | GEO

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