Project description:The proper response to shear stress involves multiple cell components of the vascular wall including endothelial cells (ECs), smooth muscle cells (SMCs), and the intercellular communications between them. Mounting evidence indicates that retinoid acid receptor-related orphan receptor-α (RORα) mediates many biological activities of melatonin including transactivation of transcriptional factors, and anti-inflammation. In this study we investigated the effect of melatonin on endothelial cells-derived exosomal miRNA expression. We used microarrays to detail the global program of gene expression underlying melatonin treatment and identified distinct classes of dys-regulated genes during this process.
Project description:Preeclampsia is a common complication of pregnancy that affects 4-5% of pregnant women around the world. At present, there is a lack of early identification of high-risk patients of preeclampsia in clinical practice, which restricts the development of disease prevention and treatment. Previous studies have indicated that plasma exosomal miRNAs in pregnant women could serve as biomarkers of preeclampsia, but few is focused on exosomal miRNAs from preeclampsia pregnancy with severe features(sPE). Therefore, we detected and compared the plasma exosomal miRNA profiles between normal pregancy and sPE to explore potential biomarkers and pathogenic mechanisms of sPE.
Project description:Exososmes, potent intercellular communicators, are supposed to contribute to metastasis formation, which we confirmed for exosomes of the metastatic rat pancreatic adenocarcinoma line BSp73ASML that promote metastatic settlement in lymph nodes and lung of poorly metastatic BSp73ASML cells with a selective CD44v4-v7 (BSp73ASML-CD44vkd) knockdown. To define the molecular pathway(s), whereby exosomes contribute to premetastatic niche preparation, we profiled mRNA miRNA of BSp73ASMLwt and BSp73ASML-CD44vkd- exosomes and evaluated the impact on potential target cells. BSp73ASML exosomes are recovered in the draining lymph node after subcutaneous injection. In vitro, they preferentially bind and are taken-up by lymph node stroma cells (LnStr) and lung fibroblasts (LuFb) that were chosen as exosome targets. BSp73ASMLwt and BSp73ASML-CD44kd exosomes contain a restricted repertoire of mRNA and miRNA, hwere the lattter differe significantly between the two lines and even more pronounced, exosomes derived thereof with a not yet explored dominance of tumor-suppressor miRNA in ASML-CD44kd cells and exosomes. Both, exosomal mRNA and miRNA are recovered in target cells and exosome-uptake is accompanied by significant changes in gene expression. We didn't observe a correlation between exosomal mRNA and changes in target cell mRNA or proteins. Instead transferred miRNA significantly affected target cell mRNA translation as demonstrated for selected, most abundant ASML exosomal miRNA besides others, miR-494 known target MAL (myelin and lymphocytes protein)/cadherin17, and miR-542-3p which targets TRAF/cadherin17. Furthermore, MMP transcription suggested to accompany cadherin17 dwon-regulation was upregulated in miR-494 or miR542-3p transfected or exosome co-cultured LnStr. Taken together, tumor exosomes target in vivo non-transformed cells in premetastatic organs. Exosome uptake induced altered target celll gene expression is strongly promoted by exosomal miRNA where we demonstrate for the first time that exosomes/exosomal miRNA from a metastasizing tumor line can modulate stroma cells from premetastatic organs. Endothelial cells lines were treated with pancreatic adenocarcinoma (AS) derived exosomes or pancreatic adenocarcinoma derived exosomes expressing tetraspanin 8. Total RNA was isolated and used to perform the Agilent gene expression microarrays. In this assay a replicate of endothelial cell lines treated with ASTspan8 were also included. Moreover, total RNA from both base line expression of endothelial cells and rat endothelial fibroblasts were also used to perfrom gene expression microarrays. RNA isolated from Rat endothelial fibroblasts treated with the exosomes derived from rat pancreatic adenocarcinoma and exosomes derived from rat pancreatic adenocarcinoma expressing tetraspanin8 were individually used to perfrom gene expression microarrays. RNA isolated from exosomes derived from rat pancreatic adenocarcinoma cell lines expressing tetraspanin were used to peform gene expresiion to see the base line expression. Another replicate were also used. RNA isolated from base line or control of rat pancreatic adenocarcinoma wild type cells and also base line RNA isolated from rat pancreatic adenocarcinoma cells lines where CD44 was knock-down.
Project description:To investigate the exosomal miRNA changes under LPS treatment in RAW 264.7 cells, 2 μg/mL LPS were added into complete medium to incubate RAW 264.7 cells. And then The exosomes were isolated and tested the exosomal miRNAs change using microarray.
Project description:To investigate whether IDO1 participated in OS malignant progression through miRNA, we selected non-metastatic OS cell line (MG63), metastatic OS cell line (143B) and non-tumorigenic immortalized osteoblastic hFOB1.19 cell line as the research object. Over Expression cell lines are contructed and exosomal miRNAs from both control and OE cell lines are sequenced to find out the differences in expression in the exosomes secreted from IDO1 overexpressing and WT cells.
Project description:We identified exosomal miRNA biomarkers for pancreatic cancer diagnostics by isolating exosomes using a recently developed magnetic nanopore isolation technology and small RNA sequencing.
Project description:Urinary exosomal miRNA profiling was conducted in urinary exosomes obtained from 8 healthy controls (C), 8 patients with type II diabetes (T2D) and 8 patients with type II diabetic nephropathy (DN) using Agilent´s miRNA microarrays.
Project description:Recently, exosome has been treated as a key mechanism for cell-to-cell communication. We thus demonstrated the roles of exosomes for the encoding messages between primary tumors and metastases. To investigate the difference between pTDE miRNA (primary tumor-derived exosomal miRNA) and mDE miRNA (metastases-derived exosomal miRNA), we isolated exosomal miRNA from each cell lines. Next, we performed small RNA sequencing for miRNA profiling. Compared with pTDE and mDE miRNAs, miR-1 is specifically abundunt in pTDE and also shows therapeutic effects to repress the growth of metastases after primary tumor resection.