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Transcriptome analysis of Wnt3a-treated triple-negative breast cancer cell lines.

ABSTRACT: We analyzed the transcriptome of two different triple negative breast cancer (TNBC) cell lines to define a comprehensive list of Wnt target genes. Cells were treated with Wnt3a for 6h, 12h or 24h. We found up-regulated and down-regulated genes in response to Wnt3a treatment. They are involved in the Wnt pathway itself, and also in TGFß, p53 and Hedgehog pathways. Thorough characterization of these novel potential Wnt target genes may reveal new regulators of the canonical Wnt pathway. The comparison of our list of Wnt target genes with those published in other cellular contexts confirms the notion that Wnt target genes are tissue-, cell line- and treatment-specific.

ORGANISM(S): Homo sapiens

PROVIDER: GSE65238 | GEO | 2015/01/24

SECONDARY ACCESSION(S): PRJNA273488

REPOSITORIES: GEO

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Transcriptome analysis of Wnt3a-treated triple-negative breast cancer cell lines.

Project description:We analyzed the transcriptome of two different triple negative breast cancer (TNBC) cell lines to define a comprehensive list of Wnt target genes. Cells were treated with Wnt3a for 6h, 12h or 24h. We found up-regulated and down-regulated genes in response to Wnt3a treatment. They are involved in the Wnt pathway itself, and also in TGFM-CM-^_, p53 and Hedgehog pathways. Thorough characterization of these novel potential Wnt target genes may reveal new regulators of the canonical Wnt pathway. The comparison of our list of Wnt target genes with those published in other cellular contexts confirms the notion that Wnt target genes are tissue-, cell line- and treatment-specific. Cells were seeded in six-well plates, serum starved overnight then treated with Wnt3a for the indicated times (6, 12 and 24 hours). Triplicates for each condition were included in the experiment.

2015-01-24 | E-GEOD-65238 | biostudies-arrayexpress

Expression data from human mesenchymal stem cells treated with Wnt3a

Project description:Wnt signaling is upregulated frequently in several cancers, including sarcomas. Since, there is cell-context dependent variation in the target gene expression, to identify canonical Wnt targets in sarcomas, we used human mesenchymal stem cells. Human mesenchymal stem cells were treated with 100ng/ml recombinant Wnt3a either 6 hrs or 24 hrs. Total RNA was extracted from untreated and Wnt3a treated samples using Qiagen's RNA extraction kit.

2011-02-16 | E-GEOD-27313 | biostudies-arrayexpress

Microarray of 3D cultured basal cells in the presence of Wnt3A

2014-10-22 | GSE55117 | GEO

Wnt3a-Activin A Synergy Induces eRNAPII Pause-Release and Counteracts a Yap1 Elongation Block during hESC Differentiation

Project description:The Wnt3a/?-catenin and Activin/Smad2,3 signaling pathways synergize to induce endodermal differentiation of human embryonic stem cells, however the mechanism is not well-understood. Using ChIP-seq and GRO-seq analyses, we report here that hESC enhancers, including Wnt3a/LEF-1 sites, hold enhancer RNAPII complexes (eRNAPII) containing high levels of Ser5P and low Ser7P. In Wnt3a signaling, ?-catenin recruits cohesin to the LEF-1:eRNAPII sites to induce enhancer-promoter looping and activate transcription of mesoendodermal (ME) genes. However, paused Ser5P-RNAPII complexes accumulate at these genes, indicating that elongation remains limiting. Subsequent Activin/Smad2,3 signaling increases P-TEFb occupancy, CTD-Ser7P, and productive elongation at ME genes. Additionally, ME genes, including EOMES and MIXL1, are repressed by the Hippo regulator, Yap1, an essential pluripotency factor. GRO-seq experiments indicate that Yap1 blocks nascent transcription and controls NELF occupancy on ME genes. Thus, Wnt3a/?-catenin and Activin/Smad2,3 pathways up-regulate transcription initiation and elongation, respectively, to overcome Yap1 repression during early hESC differentiation ChIP-seq and GROseq experiments in H1 hESCs. Cells were treated with Wnt3a (200ng/ml), Activin A (100ng/ml) or Wnt3a+Activin A (W200ng/ml+A100ng/ml) for 4h (ChIP-seq) or 6h (GRO-seq). GRO-seq in YAP depleted cells were carried out following transfection with control or YAP siRNAs . After 48h transfection, cells were left untreated or treated with Wnt3a+Activin (W200ng/ml+A100ng/ml) for additional 6h.

2015-05-04 | E-GEOD-64758 | biostudies-arrayexpress

Microarray of 3D cultured basal cells in the presence of Wnt3A

Project description:Adult stem cells have the ability to self-renew and to generate specialized cells. Self-renewal is dependent on extrinsic niche factors but few of those signals have been identified. We show that adult mammary glands contain a Wnt-responsive cell population that is enriched for stem cells. In cell culture experiments, exposure to purified Wnt protein clonally expands mammary stem cells for many generations and maintains their ability to generate functional glands in transplantation assays. We propose here that Wnt3A treated mammary stem cells retain their stemness through the regulation of its downstream target genes. We used microarrays to detail the global gene expression pattern underlying mammary stem cells response towards Wnt3A treatment and identified distinct classes of genes during this process Mammary glands from 8- to 12-week-old virgin female mice were isolated and single-cell suspension was obtained. Mammary stem cell enriched population (Lin-, CD24+, CD29hi) cells were isolated using BD FACSAria. The purity of sorted population was routinely checked and ensured to be more than 95%.Total RNA from 2nd colonies passage cultured in the presence of vehicle and Wnt3A was extracted with PicoPure (Arcturs) in accordance with the manufacturerM-bM-^@M-^Ys protocol. RNA concentration was determined with NanoDrop ND-1000, and quality was determined using the RNA 6000 Nano assay on the Agilent 2100 Bioanalyzer (Agilent Technologies). Affymetrix microarray analysis, fragmentation of RNA, labelling, hybridization to Mouse Genome 430 2.0 microarrays, and scanning were performed in accordance with the manufacturerM-bM-^@M-^Ys protocol (Affymetrix). Total 4 samples representing two sets of replicates were analyzed.

2014-10-22 | E-GEOD-55117 | biostudies-arrayexpress

Transcriptome analysis of Wnt3a-treated Wild Type, Lrp5KO , Lrp6KO and Lrp5/6KO osteoblasts

Project description:Wnt signaling is a major regulator of osteoblast differentiation and function. To investigate how Wnt3a signaling regulates osteoblastic gene expression and to identify the role of Lrp5 and Lrp6 in mediating Wnt3a signaling in osteoblasts, neonatal calvarial osteoblasts isolated from C57Bl6 (WT) and osteoblasts lacking Lrp5 (Lrp5KO), Lrp6 (Lrp6KO) and, both Lrp5 and 6 (Lrp5/6KO) were treated with Wnt3a for 24 hours and gene expression changes were quantified by RNA-seq.

2017-11-07 | GSE103492 | GEO

KIAA0753 human fibroblasts treated with control media, Smoothened Agonist (SAG) or WNT3A

Project description:Human fibroblasts from a control or a patient with compound heterozygous variants in KIAA0753 treated with SAG or WNT3A to test responses within canonical Hedgehog or WNT signaling.

2021-08-24 | GSE182286 | GEO

Characterization of in vitro cultured alveolar macrophages (AM) heterogeneity following Wnt3a treatment [single-cell RNA-seq]

Project description:To understand how Wnt signaling affects AM phenotype in vitro, we treated AMs pooled from 5 WT mice with or without Wnt3a for overnight and performed single-cell RNA sequencing analysis in the cells. Dimensionality reduction analysis of combined control or Wnt3a-treated AMs revealed eight clusters based on their gene expression pattern, and control or Wnt3a-treated AMs were almost mutually exclusive as visualized in UMAP plot.

2021-05-05 | GSE164792 | GEO

Expression data from the node and primitive streak (NPS) regions from WT and Wnt3a null embryos

Project description:The goal of this project was to elucidate the target genes and transcriptional networks activated by Wnt3a during gastrulation, a complex morphogenetic process in which the embryonic germ layers are formed and the vertebrate body plan is established. To identify downstream targets of Wnt3a, transcriptional profiling was performed using Affymetrix GeneChips on both wildtype and Wnt3a null embryos at embryonic days (E) 7.75-8. In order to maximize the likelihood of identifying direct target genes of Wnt3a, RNA was isolated from the node and primitive streak (NPS) region, an area where Wnt3a is highly expressed and at a time 12-18 hours before the morpholical phenotype of Wnt3a mutants becomes apparent.

2011-06-30 | E-GEOD-29995 | biostudies-arrayexpress

Identification of genes affected by Wnt3a in primary mouse myoblasts

Project description:In order to gain insight into the molecular events operating downstream of canonical wnt-signaling in myoblasts, we compared by microarray analysis the transcriptome of myoblast cultured for 4 hours in the presence and absence of Wnt3a.

2015-01-06 | GSE50238 | GEO

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