Identifying microRNAs relating to morphine response in BE(2)-C cell line by microRNA profiling

چکیده مقاله

MicroRNAs (miRNAs) serve as critical modulators of post transcriptional gene regulation in response to stimuli such as narcotics. Hence, the analysis of miRNA expression may elucidate the molecular mechanisms underlying cellular response to morphine. Here, the expression profile of 750 known miRNAs was generated in BE (2)-C neuroblastoma cell line by using the miRCURY LNA microRNA qPCR array system, in order to identify morphine responsive miRNAs. This population of miRNAs can be used potentially to provide an insight into the functional mechanisms of this drug. Bioinformatics analysis was used to predict the target genes of morphine responsive miRNAs, and the altered expression of selected targets was investigated through quantitative polymerase chain reaction. The result showed that a total of 10 miRNAs were up-regulated, while 33 miRNAs were down-regulated in response to morphine treatment, considering at least 1.75 fold change in expression between treated and untreated cells. Among them, hsa-miR-29a-5p, has-miR-646, has-miR-1539, has-miR-412 and has-miR-937 showed the largest changes in expression level. Furthermore, the effect of multiple morphine responsive miRNAs on one target gene (cooperativity) was considered as the criterion to select target genes for the validation analysis. Cnr1, Jazf1, Nr2c2, Lonrf2, Bnc2, C1orf21 and eight more genes were selected based on cooperativity, and validated by using real time polymerase chain reaction.