Effect of overexpression and depletion of CCDC137 on miRNA expression in hepatocellular carcinoma cells [miRNA-seq]
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ABSTRACT: To investigate the function of CCDC137 in the regulation of miRNA expression, we established CCDC137-overexpressing and CCDC137-knockdown cells.We then performed miRNA expression profiling analysis using data obtained from miRNA-seq of CCDC137-overexpressing and CCDC137-knockdown cells.
Project description:To investigate the function of CCDC137 in the regulation of gene expression, we established CCDC137-overexpressing and CCDC137-knockdown cells.We then performed gene expression profiling analysis using data obtained from RNA-seq of CCDC137-overexpressing and CCDC137-knockdown cells.
Project description:The 22Rv1 and PC-3 cells were transduced with shMIMIC human lentiviral vectors (Horizon Discovery, Cambridge, UK) to stably overexpress miRNA-23c or -4328. The SMARTvector Non-Targeting Control (NTC) was expressed to serve as a negative control. The vectors contained a turbo green fluorescent protein (turboGFP) and a puromycin resistance gene cassette. After transduction, cells were cultured in a medium supplemented with 5 µg/mL puromycin (Takara Bio, Tokyo, Japan) for antibiotic selection. Overexpression was confirmed by monitoring the turboGFP by fluorescence microscopy and by RT-qPCR analysis of miRNA-23c and -4328 levels. Relative protein quantification was performed to compare protein expression in 22Rv1 and PC-3 single cell clones overexpressing miRNA-23c and -4328, compared to corresponding NTC cells. Single cell clones overexpressing either miRNA-23c (n = 3), -4328 (n = 3) or NTC (n = 3) were analyzed in triplicate.
Project description:Analysis of Huh7 hepatocellular carcinoma (HCC) cells depleted for potassium channel KCa3.1, the intermediate conductance calcium-activated potassium channel. Result provide insight into the role of KCa3.1 in the carcinogenesis of HCC. We used microarrays to detail the gene expression differences between KCa3.1 knockdown and negative control in Huh7 HCC cells.
Project description:The incidence of TP53 loss-of-function in hepatocellular carcinoma is very high. In order to clarify the gene expression differences induced by the changes of TP53 gene, we used two human hepatocellular carcinoma cell lines, SK-HEP-1 and Hep 3B with TP53 knockdown or overexpression for RNA sequencing . SK-HEP-1 is a TP53 wild-type hepatocellular carcinoma cell line. Thus, we knockdown TP53 in SK-HEP-1. Hep 3B is a TP53 loss-of-function hepatocellular carcinoma cell line. Thus, we overexpress TP53 in Hep 3B. Results of RNA-seq analysis showed the differences after knocking-down or overexpressing TP53.
Project description:Analysis of DDX20 knockdown - hepatocellular carcinoma cells. The expression levels of genes driven by NF-kB and related with carcinogenesis, were significantly enhanced in DDX20-knockdown cells. One condition experiment, HepG2 vs. HepG2-DDX20 knockdown cells
Project description:Analysis of DDX20 knockdown - hepatocellular carcinoma cells. The expression levels of genes driven by NF-kB and related with carcinogenesis, were significantly enhanced in DDX20-knockdown cells.
Project description: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:Global miRNAs expression profilling of Caco2 cells after shRNA-mediated knockdown of PLEKHA7 Caco2 cells were infected and selected either with a non-target (NT) or with a PLEKHA7-targeting shRNA (shPLEKHA7). Total RNA was isolated from three independent sets of infections and was used to assess total miRNA expression using NanoString's nCounter Human v2 miRNA Expression Assay Kit