Unknown,Transcriptomics,Genomics,Proteomics

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Expression profiling of immortalized mature, murine T cells

ABSTRACT: The RNA expression profile of retrovirally immortalized mature, murine T cells was compared to the expression profile of mature naive, murine T lymphocytes and/or murine thymocytes of healthy C57BL/6 mice. Retroviral insertional mutagenesis can contribute to immortalization of mature T lymphocytes, Newrzela et al. 2011 Mature T lymhpocytes were retrovirally immortalized. To compare the expression profile of immortalized T cells with non-manipulated wild type cells: RNA was isolated from fresh murine T lymphocytes, thymocytes and the immortalized T-cell population (each 1x10E7 cells). For naive T lymphocytes and thymocytes we used 8 weeks old C57BL/6 wild type (WT) donor animals. Study design: Prof. Dorothee von Laer. Study design and sample preparation: Dr.Sebastian Newrzela Gene expression profiling was performed as a commercial service by Miltenyi Biotec MACS molecular, Micorarray Service/ Cologne, Germany. Responsible scientist: Alena Fitz, Silvia Rüberg. Performed: March 2008

ORGANISM(S): Mus musculus

SUBMITTER: Sebastian Newrzela

PROVIDER: E-GEOD-30349 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Disseminated tumor cells (DTCs) in the bone marrow can be detected in patients with solid tumors early on in disease progression. Via interaction with mesenchymal stromal cells (MSCs) these tumor cells may interfere with hematopoiesis. Using appropriate co-culture models, we investigated whether DTCs can change the bone marrow microenvironment by modulating MSC function with a special emphasis on their chemoattractive activity towards hematopoietic stem and progenitor cells (HSPCs). Human bone marrow derived MSCs as well as an immortalised MSC line (SCP-1) were co-cultured with MCF-7, MDA-MB231 breast carcinoma or MCF-10A non-malignant breast epithelial cells or their conditioned medium. Gene expression analysis of SCP-1 cells cultured with MCF-7 conditioned medium revealed SDF-1/CXCL12 as one of the significantly downregulated genes. Both tumor cell lines caused an inhibited SDF-1 promoter activity in SCP-1 cells, whereby MCF-7 medium decreased it to 77% and MDA-MB231 to 47%. Moreover, the SDF-1 mRNA and protein levels were significantly reduced. As a functional consequence of lower SDF-1 levels, we detected a decreased trans-well migration potential of CD34+ HSPC to MSC/tumor cell co-cultures or conditioned medium. The specificity of this chemokine mediated effect was confirmed by blocking studies with the CXCR4 antagonist AMD3100. Downregulation of SP1 transcription factor and increased miR23a levels in MSCs after contact with tumor cell medium as well as an enhanced TGFb1 expression were identified as potential molecular regulators of SDF-1 activity in MSCs. We propose an additional mechanism by which tumor cells affect the niche environment of HSPCs and therefore negatively impact hematopoiesis. Gene expression of human immortalized mesenchymal stromal cells (SCP-1) was investigated after incubation with conditioned medium of the breast carcinoma cell line MCF-7 for 24, 48, and 72 hours. Three independent experiments were performed at each time.

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Expression profiling of immortalized mature, murine T cells

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