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- W2133451685 abstract "Glioblastoma multiforme (GBM) is the most common brain tumor. Tumor microvesicles are membrane-bound outpouchings or “blebs” released from cells that are involved in intercellular communication and have been shown to transport biologically active protein and RNA species. Given the important role(s) that tumor microvesicles play in cellular communication, it is important to understand how radiation alters their release and content. We hypothesized that GBM cells stressed with ionizing radiation (IR) would release microvesicles with biological contents unique to the cellular radiation response. Microvesicles were isolated from the GBAM1 glioma stem cell line through differential centrifugations, visualized by scanning electron microscopy, and quantified based on protein concentration. Western blots, flow cytometry, and antibody arrays were used to assess protein expression. cDNA microarrays were used to assess gene expression. Differentially expressed genes were organized into gene ontology (GO) pathways, and further analyzed using the Ingenuity Pathway Analysis (IPA) software to determine relevant cellular pathways. We show that GBAM1 microvesicles retain the stem cell marker CD133. In addition, we observe an increase in the number of microvesicles released into the conditioned media after cells were treated with 4 Gy of IR. Microvesicles isolated from irradiated cells compared to microvesicles from control cells showed 397 differentially expressed genes with upregulated GO pathways related to DNA damage response, defense response, adhesion and regulation of cell death. Using IPA, the three most statistically significant molecular networks of the differentially expressed genes were (a) Cell Cycle, Cell-To-Cell Signaling and Interaction, Cellular Growth and Proliferation, (b) Cellular Movement, Inflammatory Disease, Respiratory Disease, and (c) Cell Morphology, Cell Death, DNA Replication, Recombination, and Repair. Merging these networks revealed numerous hub molecules related to DNA damage response and tumor progression, including ATM, BRCA1, BRCA2, BCL2, TP53 and PI3K. In addition, a protein cytokine antibody array on microvesicles isolated from irradiated cells compared to microvesicles from control cells showed an increase in numerous molecules related to tumor progression and invasion such as MMP-1, MMP-2, MMP-7, PAI-1, VEGFR2, VEGFR3, CXCL-16 and EGF. These initial results indicate that GBAM1 microvesicles retain stem cell markers and that microvesicles from irradiated cells are different from microvesicles secreted by un-irradiated cells. Studies are ongoing using functional studies to better assess how microvesicles from control and irradiated cells influence the phenotype of adjacent tumor and stromal cells." @default.
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- W2133451685 date "2010-11-01" @default.
- W2133451685 modified "2023-09-26" @default.
- W2133451685 title "The Profile of Glioma Microvesicles After Irradiation" @default.
- W2133451685 doi "https://doi.org/10.1016/j.ijrobp.2010.07.326" @default.
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