NGS-based miRNome reveals regulation in the miR-34/miR-449 cluster targetome in sinonasal cancers [miRNA-seq]
Ontology highlight
ABSTRACT: Using NGS-based miRNome, followed by AGO2-RIP-seq, the miR-34c and miR-449a and their direct targets were identified as factors involved in the development and progression of sinonasal cancers (SNCs). Both miR-34c and miR-449a were independent prognostic biomarkers and were associated with patient outcome.
Project description:Using NGS-based miRNome, followed by AGO2-RIP-seq, the miR-34c and miR-449a and their direct targets were identified as factors involved in the development and progression of sinonasal cancers (SNCs). Both miR-34c and miR-449a were independent prognostic biomarkers and were associated with patient outcome.
Project description:We used microarrays to detail the global programme of gene expression that occurs in response to miR-449 or miR-34 overexpression in proliferating HAECs. Each donors were transfected with pre-miR-Negative control, pre-miR-449a and pre-miR-449b. The donors 3 and 4 were morever transfected with pre-miRs-34 (34a, 34b-5p, 34c-5p).
Project description:In this study, we comparatively analyzed the members of the miR-449 family (miR-449a, miR-449b, and miR-449c) with regards to their target genes and functional effects in hepatocellular carcinoma (HCC). Microarray analysis after transient transfection of miR-449a, miR-449b, and/or miR-449c in the HCC cell line HLE identified putative target genes of miR-449a, miR-449b, and miR-449c.
Project description:In this study, we comparatively analyzed the members of the miR-449 family (miR-449a, miR-449b, and miR-449c) with regards to their target genes and functional effects in hepatocellular carcinoma (HCC). Microarray analysis after transient transfection of miR-449a, miR-449b, and/or miR-449c in the HCC cell line HLE identified putative target genes of miR-449a, miR-449b, and miR-449c. For transient overexpression of miRNAs, the HCC cell line HLE was transfected with 50 nM Allstars Negative Control or miScript miRNA mimics. MiRNA mimics for miR-449a, miR-449b, and miR-449c were either single transfected or cotransfected in equimolar amounts (16.7 nM each). Global mRNA expression profiling was performed utilizing pooled RNA of three biological replicates per microarray and two microarrays per condition.
Project description:Epithelial, non-glandular sinonasal cancers (SNCs) is a rare disease, with a global dismal prognosis. There are no recognized targeted treatments and the knowledge of molecular mechanisms involved in the resistance to available therapies is limited. Dissecting the heterogeneity of paranasal sinus cancersSNCs and providing valuable information on the biology of the malignancy is eagerly needed to improve therapeutic approaches.
Project description:The miR-34 family of microRNAs consisting of miR-34a, miR-34b and miR-34c are tumour suppressors. The annotated human miR-34b-5p has one additional base at the 5’ end of the common miR-34 family seed sequence, compared to miR-34a-5p and miR-34c-5p. This extra base results in a shift of the seed sequence, which would affect the target gene repertoire and have functional consequences. During our studies of miR-34 functions, we investigated the precise sequence of mature miR-34b-5p in human cells by deep sequencing. We found that a miR-34b-5p without the extra base was the predominant form in both non-malignant and malignant cells derived from several human tissues, indicating that the miR-34b annotation is misleading. We evaluated the functional implications of the seed shift, by comparing the effect of mimics representing the alternative miR-34b-5p sequences in MDA-MB-231 cells. In contrast to the annotated miR-34b, the endogenously expressed miR-34b displayed tumour suppressive characteristics in vitro similarly to miR-34c. These data demonstrate the importance of determining the precise sequence of a mature microRNA before exploring miRNA functions.