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Gene expression profiling and localization of Hoechst-effluxing CD45- and CD45+ cells in the Embryonic Mouse Lung

ABSTRACT: Side population (SP) cells are highly enriched in stem and progenitor cells. CD45+and CD45- SP cells were found at all developmental lung stages with the highest frequency of cells present at embryonic day 17.5 (E17.5), In order to clarify the role of these cells in lung development, we used oligonucleotide microarrays to evaluate their gene expression profiles. To do this, oligonucleotide gene arrays were performed in triplicate on RNA derived from CD45+ and CD45-SP cells, and CD45+ and CD45- non-SP populations (main population; MP) isolated from the E17.5 lung. Keywords: parallel sample

ORGANISM(S): Mus musculus

PROVIDER: GSE2057 | GEO | 2004/12/11

SECONDARY ACCESSION(S): PRJNA91299

REPOSITORIES: GEO

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Project description:Pituitary adenomas cause significant endocrine and mass-related morbidity. Not much is known about mechanisms underlying pituitary tumor pathogenesis. We searched for a side population (SP) in pituitary tumor, representing cells with high efflux capacity and potentially enriching for cancer/tumor stem cells (CSC/TSC). Human pituitary adenomas contain a SP irrespective of hormonal phenotype. This adenoma SP, as well as the further purified SP (pSP) depleted from endothelial and immune cells, enriches for cells that express ‘tumor stemness’ markers and signaling pathways, including epithelial-mesenchymal transition (EMT)-linked factors. The adenomas were found to contain self-renewing sphere-forming cells, considered a property of TSC. The sphere-initiating cells were retrieved in the pSP. Because benign pituitary adenomas do not grow in vitro and failed to expand in immunodeficient mice, the pituitary tumor cell line AtT20 was further used. We identified a SP in this cell line and found it more tumorigenic than the main population (MP). Of the two EMT-regulatory pathways tested, inhibition of Cxcr4 signaling reduced EMT-linked cell motility in vitro as well as xenograft tumor growth, whereas activation of TGFβ had no effect. The tumor pSP showed upregulated expression of the pituitary stem cell marker SOX2. Pituitaries from dopamine receptor D2 knockout (Drd2-/-) mice bearing prolactinomas, contain more pSP, Sox2+ and colony-forming cells than wildtype glands. In conclusion, we detected a SP in pituitary tumors and identified TSC-associated characteristics. Our study may contribute to a better understanding of pituitary tumor pathogenesis and lead to new therapeutic targets. We determined gene expression profiles for 5 human pituitary adenoma samples (all NF-A type). The samples were obtained immediately after transsphenoidal resection (by the neurosurgeon Dr. van Loon, Division Neurosurgery, University Hospitals Leuven). Informed consent was received from the patients. Hormonal phenotype was determined by the Department of Imaging and Pathology (University Hospitals Leuven). For each sample, CD31¯/CD45¯ SP (purified SP or pSP) and pMP cells were sorted by FACS. Separate assays were run subsequently for the two fractions. As such we present data from 10 microarrays.

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Gene expression profiling and localization of Hoechst-effluxing CD45- and CD45+ cells in the Embryonic Mouse Lung

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