Differentially expressed genes in HUVEC following miRNA-146a overexpression
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ABSTRACT: We report that miRNA-146a exerts anti-proliferative effects on endothelial cells in vitro as well as in vivo. In order to obtain insights into potential target genes, we generated samples of HUVEC overexpressing miRNA-146a and RNA sequencing was performed by Novogene (Novogene Bioinformatics Technology, Beijing, China)
Project description:We report that miRNA-146a exerts anti-proliferative effects on smooth muscle cells in vitro as well as in vivo. In order to obtain insights into potential target genes, we generated samples of SMC overexpressing miRNA-146a and RNA sequencing was performed by Novogene (Novogene Bioinformatics Technology, Beijing, China)
Project description:This study was designed to explore the role of miRNA-146a (miR-146a) and its target genes in the endothelial cells. In this study we have demonstrated that lipopolysaccharide (LPS) induced upregulation of miR-146a in the human umbilical vein endothelial cells (HUVEC) and the induction was blocked by the silencing of the toll-like receptors (TLRs) adaptor molecules MyD88 and nonspecific NF-κB inhibitor BAY 11-7082. In addition, knockdown of miR-146a by transfection of the locked nucleic acid (LNA)-antimiR-146a significantly decreased the increased cell migration and tube formation induced by LPS. A combined analysis of the bioinformatics miRanda algorithms and the whole genome expression microarray of the immunoprecipitated Ago2 ribonucleoprotein complex identified 14 transcripts as the potential target genes. Subsequent transfection with miR-146a precursor pre-miR-146a into the HUVEC validated that the CARD10 was the target gene of the miR-146a both in the transcript and protein level.
Project description:A long-prevailing model has held that the “seed” region (nucleotides 2-8) of a microRNA is typically sufficient to mediate target recognition and repression. However, numerous recent studies, both within the context of defining miRNA/target pairs by direct physical association and by directly assessing this model in vivo in C. elegans have brought this model into question. To test the importance of miRNA 3' pairing in vivo, in a mammalian system, we engineered a mutant murine mir-146a allele in which the 5' half of the mature microRNA retains the sequence of the wild-type mir-146a but the 3ʹ half has been altered to be anti-complementary to the wild-type miR-146a sequence. Mice homozygous or hemizygous for this mutant allele are phenotypically indistinguishable from wild-type controls and do not recapitulate any of the immunopathology previously described for mir-146a-null mice. Our results strongly support the conclusion that 3ʹ pairing is dispensable in the context of the function of a key mammalian microRNA.
Project description:Human vein umbilical endothelial cells (HUVEC) were transfected with pre-miR control and pre-miR 146 (Ambion) in order to identify targets (direct and indirect) downregulated by miR-146a in endothelial cells. 164 transcripts were downregulated with a fold change ≥ 1.2. Total RNA was obtained from cells transfected in HUVEC with pre-miR control and pre-miR 146a. Three independent transfections were performed for each condition. 2 replicates for control re-control and 3 for pre-miR 146a were analyzed.
Project description:To investigate the biological function ALDOA in the colorectal cancer, we established HT29 cell lines in which the ALDOA gene has been knocked down by the small interfering RNA (siRNA) technique. We used a siRNA targeting ALDOA (KD) to knockdown ALDOA expression in HT29 cells. A scrambled siRNA (NC) was used as a control. Then RNA-Seq experiment was performed by Novogene Co., Ltd. (Beijing, China) to analyze gene expression changes in HT29 cells. For each sample, three independent biological replicates were performed.
Project description:MiR-146a is an important regulator of innate inflammatory responses and is also implicated in cell death and survival. Here, we identified microglia as the main cellular source of miR-146a among mouse CNS resident cells. We further characterized the phenotype of miR-146a KO microglia cells during in vivo demyelination induced by cuprizone (CPZ) and found reduced number of CD11c+ microglia in the KO compared to WT mice. Microglia were also isolated from the brain, and the proteome was analyzed by liquid chromatography mass spectrometry.
Project description:miR-146a is a known anti-inflammatory miRNA. Intringuigly, it is overexpressed in RAS-induced senescent cells which is accompanied with a rich pro-inflammatoy secretpry phenotype. We aim to study possible sponges for miR-146a.
Project description:Purpose: To investigate the influence of grafting on adipose tissues, samples were collected from the inguinal fat tissues of mice before grafting and 1 week after grafting. Methods: The RNA-seq analysis was performed by Novogene (Beijing, China). Total RNA was drawn from the tissues with TRIzol reagent (Absin, Shanghai, China). Transcriptome sequencing were performed by using the Illumina HiSeq X Ten (Novogene Bioinformatics Technology Co., Ltd., Beijing, China). Mapping of 150-bp paired-end reads to the genes was undertaken. Result: In fat tissues one week after grafting, 28145 genes were detected. And 3271 genes were found to be upregulated, while 3745 genes were downregulated (padj < 0.05 and log2(FC) > 1), when compared with that in the natural fat tissues. Conclusions: Our study firstly reported the differentially expressed genes in adipose tissues after grafting, providind a valuable reference for fat grafting studies.
Project description:Human vein umbilical endothelial cells (HUVEC) were transfected with pre-miR control and pre-miR 146 (Ambion) in order to identify targets (direct and indirect) downregulated by miR-146a in endothelial cells. 164 transcripts were downregulated with a fold change ≥ 1.2.