a heatmap for top 10 highly-expressed lncRNAs in the glioma microarray GSE50161, in which the abscissa refers to the sample numbers while the ordinate refers to the differentially expressed lncRNAs the histogram in upper right corner is the color scale, in which each rectangle indicates the expression for each sample b NCK1-AS1 expression in glioma microarray GSE35493 c NCK1-AS1 expression in TCGA-GBM predicted on GEPIA ( ) (T, tumor N, normal) d NCK1-AS1 expression in glioma and normal brain tissue samples measured using RT-qPCR, * compared to the Normal tissues, p < 0.05 E, NCK1-AS1 expression in glioma cell lines U251, SHG-441, U87 and T98 and in human glial cell line HEB measured using RT-qPCR, * compared to the HEB cells, p < 0.05 F, NCK1-AS1 expression in brain tissues and glioma tissues validated using RNA-ISH assay, * compared to the HEB cells, p < 0.05. NCK1-AS1 is highly expressed in PC tissues. Glioma Long non-coding RNA NCK1-AS1 TRIM24 Wnt/β-catenin signaling pathway microRNA-138-2-3p. Silencing of NCK1-AS1 might inhibit the progression of glioma. Our study suggested that NCK1-AS1 might elevate TRIM24 expression and further activate the Wnt/β-catenin pathway via acting as a ceRNA for miR-138-2-3p. The in vitro results were reproduced in in vivo experiments. Artificial silencing of NCK1-AS1 or up-regulation of miR-138-2-3p led to inhibited proliferation, invasion and migration but promoted cell apoptosis of U251 cells, while up-regulation of TRIM24 reversed these changes, and it activated the Wnt/β-catenin pathway. Online predictions and the integrated experiments identified that NCK1-AS1 elevated the TRIM24 expression through sponging miR-138-2-3p, and further activated the Wnt/β-catenin pathway. High expression of NCK1-AS1 was found in glioma tissues and cells, especially in U251 cells. In vivo experiments were performed as well. The activity of the Wnt/β-catenin pathway was measured. Gain- and loss-of functions of NCK1-AS1, miR-138-2-3p and TRIM24 were performed to identify their roles in the behaviors of glioma cells. The interactions among NCK1-AS1, miR-138-2-3p and TRIM24 were validated through luciferase reporter, RNA immunoprecipitation and RNA pull-down assays. NCK1-AS1 levels in glioma tissues and normal brain tissues, and in glioma cell lines and normal human glial cells were identified. Microarray analyses were performed to explore the lncRNAs/miRNAs/genes with differential expression in glioma. This study was conducted to probe the role of long noncoding RNA (lncRNA) NCK1-AS1 in glioma progression and the involved mechanisms. The competing endogenous RNA (ceRNA) networks may play key roles in cancer progression. Glioma is a common brain malignancy with high mortality.
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