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

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE39890 | GEO | 2012/11/05

SECONDARY ACCESSION(S): PRJNA171996

REPOSITORIES: GEO

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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

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

Effect of p.Cys134Trp FOXL2 variant on gene expression in COV434 granulosa cell tumor derived cells

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

expression data from pediatric primary intracranial germ cell tumor

Project description:Intracranial pediatric germ cell tumors (GCTs) have different histological differentiations, prognosis and clinical behaviors. Prognosis of patients with germinoma and mature teratoma is good, while patients with other types of GCTs, termed as nongerminomatous malignant germ cell tumors (NGMGCTs), require more extensive drug and irradiation treatment regimen. The mechanisms underlying different prognosis of various GCT subgroups remain elusive. We presented a distinct mRNA profile correlating with GCT histological differentiation and prognosis. 13 central nervous system GCT cases with different histological subtypes are subjected to transcriptome analysis. The histological subtypes are germinoma, mixed GCT of germinoma and mature teratoma , immature teratoma , mixed GCTs of NGMGCTs category, yolk sac tumor, immature teratoma, and embryonal carcinoma.

2010-03-09 | E-GEOD-19348 | biostudies-arrayexpress

microRNA expression data from pediatric primary intracranial germ cell tumor

Project description:Intracranial pediatric germ cell tumors (GCTs) have different histological differentiations, prognosis and clinical behaviors. Prognosis of patients with germinoma and mature teratoma is good, while patients with other types of GCTs, termed as nongerminomatous malignant germ cell tumors (NGMGCTs), require more extensive drug and irradiation treatment regimen. The mechanisms underlying different prognosis of various GCT subgroups remain elusive. We presented the first miRNA profile of pediatric primary intracranial GCTs. 12 central nervous system GCT cases with different histological subtypes are subjected to miRNA expression analysis. The histological subtypes are germinoma, mixed GCT of germinoma and mature teratoma , immature teratoma , mixed GCTs of NGMGCTs category, yolk sac tumor, immature teratoma, and embryonal carcinoma.

2010-05-14 | E-GEOD-19347 | biostudies-arrayexpress

Genotyping and analysis of chromosome copy number variation (CNV) from pediatric primary intracranial germ cell tumor

Project description:Intracranial pediatric germ cell tumors (GCTs) have different histological differentiations, prognosis and clinical behaviors. Prognosis of patients with germinoma and mature teratoma is good, while patients with other types of GCTs, termed as nongerminomatous malignant germ cell tumors (NGMGCTs), require more extensive drug and irradiation treatment regimen. The mechanisms underlying different prognosis of various GCT subgroups remain elusive. We presented CNVs correlating with GCTs malignancy and clinical risk. 16 central nervous system GCT cases with different histological subtypes are subjected to genotyping and CNVs analysis. The histological subtypes are germinoma, mixed GCT of germinoma and mature teratoma, immature teratoma, mixed GCTs of NGMGCTs category, yolk sac tumor, immature teratoma, and embryonal carcinoma.

2010-06-24 | E-GEOD-19349 | biostudies-arrayexpress

expression data from pediatric primary intracranial germ cell tumor

2010-02-26 | GSE19348 | GEO

Genotyping and analysis of chromosome copy number variation (CNV) from pediatric primary intracranial germ cell tumor

2010-02-26 | GSE19349 | GEO

microRNA expression data from pediatric primary intracranial germ cell tumor

2010-02-26 | GSE19347 | GEO

MicroRNA Expression data from formalin-fixed, paraffin-embedded (FFPE) granulosa cell tumour (GCT) tissue samples

Project description:Granulosa cell tumours (GCTs) of the ovary are a unique subset of ovarian stromal tumours accounting for approximately five percent of all ovarian malignancies. Based on their pathologic characteristics and clinical features, GCTs can be further divided into two distinct subtypes: adult-GCTs and juvenile-GCTs. In this study, we investigate the use of microRNAs (miRNAs) as potential biomarkers for the diagnosis and prognostication of GCTs.

2017-07-20 | GSE70026 | GEO

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