GSE114055
Transcriptomic analysis of gastric cancer cells in response to stable over-expression of circular RNA derived from AGO2 gene (circAGO2)
Argonaute 2 (AGO2) is the core component of microRNA (miRNA)-induced silencing complex, and plays a compelling role in tumorigenesis and aggressiveness. Recent studies have implicated the emerging roles of circular RNAs (circRNAs) in cancer progression. However, the roles of circRNAs in regulating the functions of AGO2 in cancer still remain elusive. We indentify one intronic circRNA derived from AGO2 gene (circAGO2) as a novel regulator of AGO2-miRNA complexes and cancer progression. To investigate the mechanisms underlying the oncogenic functions of circAGO2, we employed the Illumina HiSeq X Ten as a discovery platform to analyze the transcriptome profiling changes of human gastric cancer MKN-45 cells in response to stable over-expression of circAGO2. The results showed that stable over-expression of circAGO2 led to altered expression of 624 human mRNAs, including 257 up-regulated genes and 367 down-regulated genes. Furthermore, we validated the RNA-seq results by real-time RT-PCR with high identity. Overall, our results provided fundamental information about the transcriptomic changes in response to circAGO2 over-expression in human cancer cells, and these findings will help us understand the pathogenesis of cancer progression. Overall design: Total RNA of cells stably transfected with empty vector or circAGO2 was extracted using the TRIzol® reagent according to the manufacturer's instructions. RNA concentration was measured using a Qubit® RNA Assay Kit with a Qubit® 2.0 Fluorometer (Life Technologies, Inc.), and integrity was assessed using the RNA Nano 6000 Assay Kit with a Bioanalyzer 2100 system (Agilent Technologies, CA). Library preparation and transcriptome sequencing on an Illumina HiSeq X Ten platform were performed by Novogene Bioinformatics Technology Co., Ltd. (Beijing, China), and 100 bp paired-end reads were generated. HTSeq v0.6.0 was used to count the reads numbers mapped to each gene.