The effect of Sparstolonin B (SsnB) on gene expression in HUVECs
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE44597 | GEO | 2014/02/20
SECONDARY ACCESSION(S): PRJNA190904
REPOSITORIES: GEO
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Project description:Sparstolonin B is a novel bioactive compound isolated from Sparganium stoloniferum, an herb historically used in Traditional Chinese Medicine as an anti-tumor agent. SsnB has previously demonstrated anti-angiogenic properties. In functional assays, SsnB inhibited endothelial cell tube formation (Matrigel method) and cell migration (Transwell method) in a dose-dependent manner. We used microarrays to examine how SsnB affected the gene expression of human umbilical vein endothelial cells (HUVECs), focusing in particular on pathways related to angiogenesis. Two plates of HUVECs were exposed to 100 micromolar SsnB and two plates of HUVECs were exposed to Vehicle Control (1:1000 dilution of DMSO). After 24 hours, RNA was extracted for microarrays and gene expression was analyzed.
2014-02-20 | E-GEOD-44597 | biostudies-arrayexpress
Project description:Sparstolonin B is a novel bioactive compound isolated from Sparganium stoloniferum, an herb historically used in Traditional Chinese Medicine as an anti-tumor agent. SsnB has previously demonstrated anti-angiogenic properties. In functional assays, SsnB inhibited endothelial cell tube formation (Matrigel method) and cell migration (Transwell method) in a dose-dependent manner. We used microarrays to examine how SsnB affected the gene expression of human coronary artery endothelial cells (HCAECs), focusing in particular on pathways related to angiogenesis.
2014-02-20 | GSE44596 | GEO
Project description:Sparstolonin B is a novel bioactive compound isolated from Sparganium stoloniferum, an herb historically used in Traditional Chinese Medicine as an anti-tumor agent. SsnB has previously demonstrated anti-angiogenic properties. In functional assays, SsnB inhibited endothelial cell tube formation (Matrigel method) and cell migration (Transwell method) in a dose-dependent manner. We used microarrays to examine how SsnB affected the gene expression of human coronary artery endothelial cells (HCAECs), focusing in particular on pathways related to angiogenesis. Three plates of HCAECs were exposed to 100 micromolar SsnB and three plates of HCAECs were exposed to Vehicle Control (1:1000 dilution of DMSO). After 24 hours, RNA was extracted for microarrays and gene expression was analyzed.
2014-02-20 | E-GEOD-44596 | biostudies-arrayexpress
Project description:To identify the miRNAs specifically upregulated during tube formation of primary endothelial cells , HUVECs were cultured in culture plates with or without matrigel-coating and the miRNAs from them were compared.
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Project description:Kallikrein-related peptidase 3 (KLK3, also known as prostate-specific antigen, PSA) is a chymotrypsin-like kallikrein that has been shown to have anti-angiogenic properties. KLK3 expression in prostate cancer is associated with low microvessel density. We have previously shown in a human umbilical vein endothelial cell (HUVEC) model that the anti-angiogenic effect of KLK3 is related to its enzymatic activity. However, the mechanism of this effect remains to be clarified. To this end we have used a DNA microarray to study the KLK3-induced changes in gene expression associated with reduction of HUVEC tube formation. Among the 41000 genes studied, 311 were differentially expressed in control and KLK3-treated cells. These changes were enriched in several pathways, including the pathways associated with proteasome, ubiquitin-mediated proteolysis, focal adhesion and regulation of actin cytoskeleton. Furthermore, the changes were opposite to those described previously to occur during tubulogenesis. In conclusion, our results show that KLK3 induces gene expression changes in HUVECs. While these changes might be relevant for the mechanism by which KLK3 exerts its anti-angiogenic activity, it cannot be judged by the present results whether the changes are secondary to morphogenic differentiation of the cells or the primary mechanism mediating the effect of KLK3. Keywords: KLK3 treatment, tube formation Human umbilical vein endothelial cells (HUVECs) were isolated from umbilical cord veins and grown in Endothelial Cell Basal Medium supplemented with an Endothelial Cell Growth Medium Supplement Pack (PromoCell). The cells were cultured on Matrigel basement membrane prepara-tion together with 500 nM enzymatically active KLK3 (PSA). Phosphate buffered saline (PBS) was used for control. HUVECs were grown on Matrigel for 18 h. RNA was isolated from KLK3-treated and control HUVECs from three different individuals. Cell Recovery Solution (BD Biosciences) was used to detach cells from the Matrigel. Total RNA was isolated with the RNeasy Mini Kit (Qiagen) according to manufacturerâ??s instructions. RNA con-centration and quality of the samples was determined with an Agilent 2100 bioanalyser (Agilent Technologies) by measuring the RNA integrity number (RIN). All samples contained high quality RNA (RIN 9.5-10). DNA microarray analysis was performed with the Agilent Whole Human Genome Oligo Microar-ray 4x44K (G4112F) (Agilent Technologies Inc.) following the manufacturerâ??s protocol. Total RNA (1 ï?g) was labeled using the RNA input fluorescent linear amplification kit according to the manufacturer's recommendations (Agilent Technologies Inc.). RNA was hybridized on Agilent's Human 4x44K Oligo Microarrays containing 44000 features (41000 unique genes and transcripts). Pre-processing of the data was performed by Feature Extraction software (v 7.5.1).
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Project description:Purpose: The goal of this study is to compare endothelial transcriptome affected by OASL knockdown under basal or stimulated conditions by utilizing RNA-seq. Methods: Endothelial mRNA profilies of siCTL or siOASL transfected HUVECs were generated by illumina sequencing method, in duplicate. Following an optimized data analysis workflow, RNA sequence reads were counted using StringTie-2.1.3b and used to identify genes differentially expressed between siCTL- and siOASL-transfected HUVECs treated with or without TNFα and IFNγ. Results: DESeq2 was used to identify genes differentially expressed between siCTL- and siOASL-transfected HUVECs under basal or stimulated conditions. Genes showing <10 read counts total were pre-filtered. Conclusions: Our study represents the first analysis of endothelial mRNA affected by OASL knockdown with biologic replicates, generated by RNA-seq.
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Project description:MicroRNAs (miRNAs) are small (18-22 nucleotides), non-coding RNAs that suppress the translation of target mRNAs by binding to their 3` untranslated region, and regulate gene networks, helping to control cell function and phenotype. Recently, the existence of extracellular miRNAs enclosed in exosomes has raised the possibility that they play an important role in cell-cell communication. The goals of this study were to elucidate whether or not miRNAs secreted from neoplastic cells transfer into endothelial cells and works functionally active in the recipient cells. In the present study, we investigated the interaction of a leukemia cell line (K562) and human umbilical vein endothelial cells (HUVECs) using a non-contact co-culture system, and assess the expression profiles of intercellular and extracellular miRNAs in HUVECs and K562 cells using Taqman MicroRNA Array v2.0 (Applied Biosystems, Bedford, MA). We compared the miRNA expression profiles between 1) K562 cells or HUVECs; 2) intercellular and extracellular; 3) mono-culture medium and co-culture medium. K562 cells and HUVECs were non-contact co-cultures using Transwell (Corning, pore size: 0.45µm) for 24 hr. The culture medium was collected and filtered through a 0.45 µm PVDF filter (Millipore, Billerica, MA). The intercellular and extracellular RNAs were isolated from K562 cells, HUVECs and each culture medium using the mirVana PARIS kit (Ambion, Austin, TX). The expression profile was determined using the Human Taqman MicroRNA Arrays A (Applied Biosystems). QRT-PCR was carried out on an Applied Biosystems 7900HT thermal cycler. The thermal cycling conditions consisted of 10 min at 95°C, followed by 40 cycles at 92°C for 15 s and 60°C for 1 min.
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Project description:Endothelial cells are critical for angiogenesis, and microRNAs plays important roles in this process. We investigated the regulatory role of microRNAs in endothelial cells of hepatocellular carcinoma (HCC) by examining the microRNA expression profile of human umbilical vein endothelial cells (HUVECs) in the absence or presence of human HCC cells, and identified miR-146a as the most highly up-regulated microRNA. Furthermore, we revealed that miR-146a promoted the expression of platelet-derived growth factor receptor (PDGFRA) in HUVECs, and this process was mediated by BRCA1. Overexpression of PDGFRA in the ECs of HCC tissues was associated with microvascular invasion, and predicted a poorer prognosis. These results suggest that MiR-146a plays a key role in regulating the angiogenic activity of ECs in HCC through miR-146a-BRCA1-PDGFRA pathway. MiR-146a may emerge as a potential anti-angiogenic target on ECs for HCC therapy. We have employed whole genome OneArray to examine the genome expression changes of HUVECs overexpressing miR-146a.
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