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.
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.
Project description:Transcriptional profiling of human MCF-7 breast cancer cells comparing MCF-7 cells treated with control medium (DMEM/F12 + 0,5% BSA) with MCF-7 cells treated with conditioned medium of cancer-associated adipose tissue (CMCAAT) obtained from 2 breast cancer patients. Goal was to determine the effects of CMCAAT treatment on global MCF-7 gene expression.
Project description:Paracrine signals relayed between heterogenous cell types through small and large extracellular vesicles promote cancer cell growth, invasion and metastasis. Microplasts, also called as cytoplasts or cytokineplasts are large extracellular vesicles (0.5-11µm diameter) originating from migratory cells. Microplasts are known to be associated with invasive phenotypes of cancer cells and promote metastasis. Treatment with macrophage conditioned medium induces significant shedding of microplasts from MCF-7 breast adenocarcinoma cells. To characterise microplasts and to study their potential role in intercellular communication in cancer, we isolated microplasts derived from macrophage conditioned medium treated MCF-7 cells and investigated their protein cargo through LC-MSMS.
Project description:Here, we report a transcriptomics analysis on MCF-7 breast cancer cells exposed to adipocyte conditioned medium (CM) and focus on the functional relevance that CM-affected pathways and processes and related biomarkers may have in breast cancer response to obesity.
Project description:Transcriptional profiling of human MCF-7 breast cancer cells comparing MCF-7 cells treated with control medium (DMEM/F12 + 0,5% BSA) with MCF-7 cells treated with conditioned medium of cancer-associated adipose tissue (CMCAAT) obtained from 2 breast cancer patients. Goal was to determine the effects of CMCAAT treatment on global MCF-7 gene expression. Two-condition experiment: MCF-7 control vs. CMCAAT treated cells. Biological replicates: 3 control replicates pooled as 1 (LL 1-2-4), 4 CMCAAT replicates (LL 6, LL 8, LL 9, LL 10)
Project description:To investigate whether conditioned medium (CM) from cancer cell culture impacts dynamic changes of the transcriptome in myotubes differentiated from hMuSC, we ran RNA sequencing from differentiated hMuSC that were treated by multiple breast cancer cell lines-conditioned medium. Among 14208 readable mRNAs, MCF-7 CM treatment induced significant changes in 2680 genes, MB-468 CM caused significant changes in 2674 genes, and SKBR-3 CM resulted in significant changes in 1823 genes in myotubes from differentiated hMuSC, compared to control group, respectively. Among them, 961 genes had significant upregulation or downregulation presented in differentiated myotubes over all 3 breast cancer cell lines CMs
Project description:Proteins secreted into the conditioned medium by human monocytic cell line, THP-1, or human buffy coat-derived monocytes polarised to either an M1 (incubation for 48 h with 100 ng/mL LPS and 20 ng/mL IFN-gamma) or M2 (incubation for 48 h with 20 ng/mL IL-4) phenotype. The conditioned medium was enriched for anionic molecules (including proteoglycans) by anion exchange chromatography prior to analysis.
Project description:This study aims to establish a comprehensive, genome-scale metabolic model (GSMM) combining different omics data of different regulatory levels of breast cancer cells in order to elucidate the way that these regulatory levels affect or determine each other. Together with transcriptomics, proteomics and metabolomics data obtained, fluxomics data was also to be integrated. In order to get the fluxomics data, MCF cells were incubated with either the medium containing [1,2-13C2]-D-glucose or [U-13C5]-L-glutamine for 8 or 24 hours. A good GSMM should answer any perturbations applied; therefore, different incubation conditions should be applied to generate different metabolic profiles that will help to test if the model of breast cancer works well. To this end, MCF cells were either administered oligomycine, which is an ATP synthase inhibitor, or were deprived of glutamine. Cells were counted before and after each incubation time so as to get the cell proliferation curve. Later, at the corresponding time points, cells and media were frozen to measure the concentrations of glucose, glutamine and lactate (not shown in the file) and the mass isotopomer distributions of glutamate, ribose, lactate, amino acids and TCA cycle intermediates (shown in the file). Finally, all these data was combined in order to estimate the flux rates of various reactions within the central carbon metabolism (not shown in the file). The fluxomics data obtained in this way is to be introduced in the GSMM designed.
