0000000000189032
AUTHOR
Elīna Zandberga
Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages.
Abstract. Background Macrophages are one of the most important players in the tumor microenvironment. The polarization status of tumor associated macrophages into a pro-inflammatory type M1 or anti-inflammatory type M2 may influence cancer progression and patient survival. Extracellular vesicles (EVs) are membrane-bound vesicles containing different biomolecules that are involved in cell to cell signal transfer. Accumulating evidence suggests that cancer-derived EVs are taken up by macrophages and modulate their phenotype and cytokine profile. However, the interactions of cancer-derived EVs with monocytes and macrophages at various differentiation and polarization states are poorly understo…
Additional file 5: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
Effect of SW480 and SW620-derived EVs on biomolecule secretion patterns of monocytes and M0, M1 and M2 macrophages. Luminex data analysis showing TNFα, IL-6, CXCL10, IL-23, IL-10, MMP9, IL-1β and CCL22 concentration in cell culture supernatants of monocytes (M) and M0, M1 and M2 macrophages following incubation with SW480 and SW620 EVs or without them (control). The graphs represent mean ± SD (n = 3). (PDF 39 kb)
High expression of GLI1 is associated with better survival in advanced SCLC
Aim Aberrant Sonic hedgehog (Shh) pathway signaling has been described in small cell lung cancer (SCLC), as well discrepancies, when analyzing expression of pathway components in SCLC cell lines vs tumor biopsies. Shh key component GLI1 was evaluated in advanced SCLC and data correlated with patient survival. Materials and methods GLI1 expression was analyzed by quantitative real-time polymerase chain reaction in pre-treatment fresh frozen tumor biopsies of 12 advanced SCLC patients and mRNA level of GLI1 was compared in short-term vs long-term survivor's samples (stratified by median survival, independent samples t-test). Results Expression of GLI1 mRNA was significantly higher in long-ter…
Additional file 3: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
Effect of temperature on the SW480 EV uptake in THP-1 monocytes. Flow cytometry histograms showing Syto RNA Select fluorescence intensities of untreated (left) and Syto RNA Select-labeled SW480 EV-treated THP-1 monocytes following incubation at 4 °C (middle) and 37 °C (right). Histogram markers show the percentage of Syto RNA Select-positive cells. (PDF 53 kb)
Additional file 2: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
SW480 and SW620-derived EV effect on monocyte (M) and macrophage (M0, M1, M2) viability. a OD values at 450 nm which are in direct proportion of viable cell counts. b SW480 and SW620 EV cytotoxicity on THP-1 monocytes and M0, M1 and M2 macrophages. The graphs represent mean ± SEM (n = 3). Statistical analysis carried out with the t-test. *p ≤ 0.05, **p ≤ 0.01 vs. untreated cell control of the respective monocyte-macrophage cell subset. (PDF 50 kb)
Expression of the Sonic Hedgehog Embryonic Signalling Pathway Components in Matched Pre-Treatment and Relapsed Small Cell Lung Cancer Biopsies
Abstract Cancer stem cells may be responsible for tumour regrowth and acquisition of resistance in small cell lung cancer (SCLC). The Hedgehog pathway regulates survival and proliferation of tissue progenitor and stem cell populations, promoting the expression of stem cell related and proliferative genes. We evaluated the Sonic Hedgehog (Shh) embryonic signalling pathway in relapsed SCLC. Expression levels of Shh related genes GLI1, SMO, SUFU, PTCH1, HHIP, BCL2, BMI, ZEB1, ZEB2, N-MYC, Twist1 were analysed by qRT-PCR in matched pre-treatment and relapsed tumour fresh frozen biopsies of three SCLC patients. Expression of each gene was compared using the paired samples t-test, as well as comp…
A novel 3D heterotypic spheroid model for studying extracellular vesicle-mediated tumour and immune cell communication
Cancer-derived extracellular vesicles (EVs) have emerged as important mediators of tumour-host interactions, and they have been shown to exert various functional effects in immune cells. In most of the studies on human immune cells, EVs have been isolated from cancer cell culture medium or patients' body fluids and added to the immune cell cultures. In such a setting, the physiological relevance of the chosen EV concentration is unknown and the EV isolation method and the timing of EV administration may bias the results. In the current study we aimed to develop an experimental cell culture model to study EV-mediated effects in human T and B cells at conditions mimicking the tumour microenvi…
Additional file 4: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
TNFα, IL-23, IL-6, IL-1 β, CXCL10, CCL22, IL-10 and MMP9 secretion profile at different monocyte-macrophage differentiation stages. The graphs represent average biomolecule concentrations SEM (n = 3). Statistical analysis carried out with one-way ANOVA test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and **** ≤ 0.0001 vs. untreated cell control of the respective monocyte-macrophage cell subset. (PDF 63 kb)
Additional file 1: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
Experimental design of the THP-1 monocyte to macrophage differentiation showing the time points for the addition of EVs and stimulatory molecules. Below the experimental design, representative light microscopy images show morphology of THP-1 monocytes (M), M0 macrophages (M0), M1 macrophages (M1) and M2 macrophages (M2) (n = 4). Scale bar 100 μm. Representative flow cytometry dot plots show CD14, HLA-DR, CD206 and CD68 marker expression at M, M0, M1 and M2 stages. (JPG 1845 kb)