Molecular changes induced by melanoma cell conditioned medium (MCM) in HUVEC cells.
ABSTRACT: Malignant melanoma is a complex genetic disease and the most aggressive form of skin cancer. Melanoma progression and metastatic dissemination fundamentally relies on the process of angiogenesis. Melanomas produce an array of angiogenic modulators that mediate pathological angiogenesis. Such tumor-associated modulators arbitrate the enhanced proliferative, survival and migratory responses exhibited by endothelial cells, in the hypoxic tumor environment. The current study focuses on melanoma-induced survival of endothelial cells under hypoxic conditions. Melanoma conditioned media were capable of enabling prolonged endothelial cell survival under hypoxia, in contrast with the conditioned media derived from melanocytes, breast and pancreatic tumors. To identify the global changes in gene expression and further characterize the pro-survival pathway induced in endothelial cells, we performed microarray analysis on endothelial cells treated with melanoma conditioned medium under normoxic and hypoxic conditions. Huvec cells were grown in melanoma conditioned medium or DMEM 10% FCS for 12 h under hypoxic or normoxic conditions. In order to identify the transcripts modulated by melanoma CM, samples treated with MCM were compared to those grown in DMEM alone.
Project description:Human Umbelical Vein Endothelial Cells (HUVEC) were cultured in vitro and exposed to 1% oxygen or normoxia for 24 hours. Then, small RNA libraries were prepared and sequenced using sequencing-by-ligation technique (Solid3 Plus Sequencer platform). Alignment versus miRBase v14.0 identified >400 different microRNA/microRNA* species expressed in HUVEC. A complex repertoire of isomiR was also observed. For novel miRNA discovery, reads were mapped against the human genome. We identified 511 candidate new microRNAs and we validated 18 of them with independent techniques. 4 samples: 2 independent HUVEC populations were analyzed, each both normoxic and hypoxic
Project description:Human umbilical vein endothelial cells (HUVECs) were incubated for 48 h under normoxic or hypoxic (1% oxygen) conditions. Changes in transcript and exon levels were analyzed. Overall design: 6 samples; 2 conditions: normoxia vs. hypoxia (1% oxygen); 3 replicates per condition
Project description:Human umbilical vein endothelial cells (HUVECs) were incubated for 48 h under normoxic or hypoxic (1% oxygen) conditions. Changes in transcript and exon levels were analyzed. 6 samples; 2 conditions: normoxia vs. hypoxia (1% oxygen); 3 replicates per condition
Project description:To determine whether hypoxia augments the activity of exosomes by modulating miRNAs, we use miRNA array profile to find the differential expression of miRNAs of exosomes in different culture conditions (normoxia, hypoxia, hypoxia supplemented with GW4869 which is an inhibitor of exosomes generation). Overall design: Three samples were processed for each kind of exosome that derived from normoxic-MSCs, hypoxic-MSCs, and hypoxic-MSCs suppled with GW4869 (N-EXO, H-EXO, H+G-EXO). H-Exosome was isolated from conditioned medium of MSCs under hypoxia condtion 24h. The fragmentation mixtures were hybridized to an Agilent- Mouse microRNA array 21.0
Project description:To define ncRNA expression in hypoxic endothelial cells, we applied pro-angiogenic hypoxia to cultured endothelial cells. Afterwards total RNA was isolated and underwent genechip analysis. HUVECs were subjected to normoxic or hypoxic (0.1-0.2% O2) cell culture conditions.
Project description:To define ncRNA expression in hypoxic endothelial cells, we applied pro-angiogenic hypoxia to cultured endothelial cells. Afterwards, total RNA was isolated and underwent RNA-seq analysis. HUVECs were subjected to normoxic or hypoxic (0.1-0.2% O2) cell culture conditions.
Project description:Tubular endothelial cells were cultured in hypoxic conditions (1% O2) and treated or not with microvesicles derived from endothelial progenitor cells.<br>mRNA profiling of hypoxic cells, treated or not with MVs, was analyzed after nromalization with mRNA profiling of normoxic cells.<br><br>This experiment was updated on 27th Jan 2011 to correct descriptions.
Project description:Evaluation of microRNA expression profile of microvesicles (MVs) derived from endothelial progenitor cells (EPCs) cultured in different oxygen concentrations (normoxic/hypoxic conditions)
Project description:Outcome prediction classifiers were successfully constructed through expression profiling of a total of 1,329 miRNAs in MKN1, gastric cancer cell line under normoxic and hypoxic conditions. In the study presented here, MKN1 under normoxic and hypoxic conditions was used to acquire expression profiles of a total 1,329 unique miRNAs.