Effect of depletion of FOXL2 on gene expression in KGN cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE227040 | GEO | 2023/11/06
REPOSITORIES: GEO
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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
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
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.
2012-11-05 | GSE39890 | GEO
Project description:In order to identify novel FOXL2 targets, we have transfected KGN cells using an expression vector containing the coding sequence of FOXL2 or, as a reference, the empty vector (mock transfection). The transcriptome perturbation induced by FOXL2 overexpression was then followed by DNA chips (i.e. FOXL2- vs. mock-transfected cells) using the platform developed by NimbleGen Systems, Inc.
2007-03-02 | E-MEXP-985 | biostudies-arrayexpress
Project description:To investigate the dysfunction of FOXL2-C134W in human granulosa cells, we established homozygous cell lines for FOXL2 WT by CRISPR/Cas9 technologies.
2024-02-22 | GSE225781 | GEO
Project description:FOXL2 is a transcription factor essential for female fertility, expressed in somatic cells of the ovary, notably granulosa cells. In the mouse, Foxl2 deletion leads to partial sex reversal postnatally. However, deletion of the gene in 8-week-old females leads to granulosa to Sertoli cell transdifferentiation. We hypothesised that different outcomes of Foxl2 deletion in embryonic versus adult ovary may depend on a different role played across ovarian development. Therefore, we characterised the dynamics of gene expression and chromatin accessibility changes in purified murine granulosa cells across key developmental stages (E14.5, 1 and 8 weeks). We then performed genome-wide identification of FOXL2 target genes and on-chromatin interacting partners by ChIP-SICAP. We found that FOXL2 regulates more genes at postnatal stages, through the interaction with factors regulating primordial follicle activation (PFA), such as NR5A2, and others regulating steroidogenesis including AR and ESR2. As a proof of principle experiment, we chose one FOXL2 interactor, Ubiquitin specific protease 7 (USP7) and showed that deletion of this gene in granulosa cells leads to a blockage of PFA, impaired ovary development and sterility. Our study constitutes a comprehensive resource for exploration of the molecular mechanisms of ovarian development and causes of female infertility.
2024-03-25 | PXD043951 | Pride
Project description:The Foxl2 transcription factor is required for ovarian function during follicular development. Our approach to begin to understand Foxl2 function is through the identification of Foxl2 regulated genes in the ovary. Transiently transfected KK1 mouse granulosa cells were used to identify genes that are potentially regulated by Foxl2. KK1 cells were transfected in three groups (mock, activated, and repressed) and twenty-four hours later RNA was isolated and submitted for Affymetrix microarray analysis. Experiment Overall Design: To increase the potential of Foxl2 to alter gene expression levels of putative target genes, two fusions were constructed consisting of Foxl2 fused to the activation domain of the Herpes simplex virus VP16 transcription factor (Foxl2-VP16) and Foxl2 fused to the repression domain of the murine MAD transcription factor (Foxl2-MAD). Levels of gene expression were compared between mock transfected cells and those that were transfected with Foxl2-VP16 and Foxl2-MAD, respectively.
2009-11-15 | E-GEOD-18891 | biostudies-arrayexpress
Project description:It has previously been shown that FOXL2 and ESR1 cooperate to repress the testis-determining gene Sox9 in murine granulosa cells, and suggested that FOXL2/ESR1 cooperation may be central to granulosa cell differentiation (Uhlenhaut et al., 2009). However, no study has so far compared the DNA-binding of FOXL2 and ESR1 at the genomic level or analyzed the impact of FOXL2 on ESR1 binding to its regulatory elements. Here, we have analyzed and compared the genomic locations recognized by ESR1 and FOXL2 in E2-treated primary murine granulosa cells. Input DNA, FOXL2 and ESR1 ChIP
2015-07-01 | E-GEOD-62545 | biostudies-arrayexpress
Project description:In order to identify genes misregulated by the adult OGCT-associated FOXL2 p.C134ZW mutant in granulosa cells, we have transfected COV434 cells (negative for the mutation) using an expression vector containing the wildtype or mutant coding sequence of FOXL2 or, as a reference, the empty vector pcDNA. The transcriptome perturbation induced by FOXL2 overexpression was then followed by DNA microarray using a platform developed by NimbleGen Systems, Inc.
2013-03-06 | E-MTAB-1017 | biostudies-arrayexpress
Project description:It has previously been shown that FOXL2 and ESR1 cooperate to repress the testis-determining gene Sox9 in murine granulosa cells, and suggested that FOXL2/ESR1 cooperation may be central to granulosa cell differentiation (Uhlenhaut et al., 2009). However, no study has so far compared the DNA-binding of FOXL2 and ESR1 at the genomic level or analyzed the impact of FOXL2 on ESR1 binding to its regulatory elements. Here, we have analyzed and compared the genomic locations recognized by ESR1 and FOXL2 in E2-treated primary murine granulosa cells.
2015-07-01 | GSE62545 | GEO