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Differential expression of genes after alterations in cellular levels of hsa-miR-195

ABSTRACT: The experiment was designed to identify the genes which get altered after over-expression and depletion of hsa-miR-195 levels in MCF7 and HEK 293T cells. Recent research has established that microRNAs (miRNAs) are key regulators of many cell processes including those often deregulated in cancer such as apoptosis. In our previous work we have shown that hsa-miR-195 targets the key anti-apoptotic molecule BCL2 and its over-expression induces apoptosis and augments sensitivity of cancer cells towards chemotherapeutic drug. These results suggested a possible anticancer role of miR-195 and therefore oblige its complete functional characterization. In this study global effect of alterations in cellular miR-195 levels on transcriptome was analysed in order to identify the regulatory functions of miR-195. Together our findings highlight a novel and important role of hsa-miR-195 potentially opening new avenues for the treatment of various diseases.

ORGANISM(S): Homo sapiens

PROVIDER: GSE43656 | GEO | 2016/02/01

SECONDARY ACCESSION(S): PRJNA186940

REPOSITORIES: GEO

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Project description:Specificity of interaction between a microRNA (miRNA) and its targets crucially depends on the seed region located in its 5’-end. It is often implicitly considered that two miRNAs sharing the same biological activity should display similarity beyond the strict six nucleotide region that forms the seed, in order to form specific complexes with the same mRNA targets. We have found that expression of hsa-miR-147b and hsa-miR-210, though triggered by different stimuli (i.e. lipopolysaccharides and hypoxia, respectively), induce very similar cellular effects in term of proliferation, migration and apoptosis. Hsa-miR-147b only shares a “minimal” 6-nucleotides seed sequence with hsa-miR-210, but is identical with hsa-miR-147a over 20 nucleotides, except for one base located in the seed region. Phenotypic changes induced after heterologous expression of miR-147a strikingly differ from those induced by miR-147b or miR-210. In particular, miR-147a behaves as a potent inhibitor of cell proliferation and migration. These data fit well with the gene expression profiles observed for miR-147b and miR-210, which are very similar, and the gene expression profile of miR-147a, which is distinct from the two others. Bioinformatics analysis of all human miRNA sequences indicates multiple cases of miRNAs from distinct families exhibiting the same kind of similarity that would need to be further characterized in terms of putative functional redundancy. Besides, it implies that functional impact of some miRNAs can be masked by robust expression of miRNAs belonging to distinct families. To compare the set of transcripts targeted by hsa-miR-147a, hsa-miR-147b and hsa-miR-210, we overexpressed these miRNAs in human lung adenocarcinoma A549 cells by transfecting them with synthetic pre-miRNAs or a synthetic “negative” pre-miRNA as a control (miR-Neg). RNA samples were harvested at 48 hours post-transfection and 3 independent experiments were carried out. 48 hours post-transfection, 3 independent experiments were performed in dye-swap: hsa-miR-147a versus miR-Neg; hsa-miR-147b versus miR-Neg; hsa-miR-210 versus miR-Neg.

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Differential expression of genes after alterations in cellular levels of hsa-miR-195

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