Elucidating the microRNA-203 specific biological processes in glioblastoma cells from comprehensive high-throughput RNA-sequencing transcriptome profiling

We report the application of high-throughput RNA-sequencing Profile of miR-203 expressing glioma U251 cells. Glioblastoma (GBM) is the most common primary malignant intracranial adult brain tumor. Allelic deletion on chromosome 14q plays an essential role in GBM pathogenesis, and this chromosome 14q site was thought to harbor multiple tumor suppressor genes associated with GBM, a region that also encodes microRNA-203 (miR-203). This study was conducted to identify whole transcriptome profile changes associated with miR-203 expression by high-throughput NGS-RNA sequencing. Total RNA samples are quantified using Nanodrop and qualified by agarose gel electrophoresis. The mRNA is enriched by oligo(dT) magnetic beads or the total RNA is depleted of rRNAs by Arraystar rRNA Removal Kit if the RNA sample is degraded or is of prokaryotic origin. We use Illumina kits for the RNA-seq library preparation, which include procedures of RNA fragmentation, random hexamer primed first strand cDNA synthesis, dUTP based second strand cDNA synthesis, end-repairing, A-tailing, adaptor ligation and library PCR amplification. Finally, the prepared RNA-seq libraries are qualified using Agilent 2100 Bioanalyzer and quantified by qPCR absolute quantification method. The sequencing is performed using Illumina Hiseq 4000. Overall design: Examination of miR-203 specific biological processes in U251 cells using high-througput RNA-sequencing transcriptome profiling