Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression signatures in a human bladder cancer cell line SW780 transfected with miR-137 precursor molecule

ABSTRACT: To clarify the effect of miRNAs, we carried out a gene expression microarray analysis of SW780 cells transfected with a miR-137 precursor or a negative control. We found that 1,326 probe sets (1,016 unique genes) were downregulated (>2-fold) by ectopic miR-137 expression, including the previously reported miR-137 target genes CDK6, CDC42 and AURKA. Moreover, Gene Ontology analysis revealed that genes related to the cell cycle were significantly enriched among the affected genes. SW780 cells were transfected with a Pre-miR-137 miRNA Precursor Molecule (Ambion) or Pre-miR miRNA Molecules Negative Control #1 (Ambion). Forty-eight hours after transfection, total RNA extraction was carried out, and gene expression signatures were analyzed.

ORGANISM(S): Homo sapiens

SUBMITTER: Hiromu Suzuki

PROVIDER: E-GEOD-41760 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Hemopoiesis entails a series of hierarchically organized events that proceed throughout cell specification and terminates with cell differentiation. Commitment needs the transcription factors effort that, in concert with microRNAs, drives cell fate specification, answering to promiscuous patterns of gene expression by turning on lineage-specific genes and repressing alternate lineage transcripts. Therefore microRNAs and mRNAs cooperate to direct cell fate decisions. We obtained microRNAs profiles from human CD34+ hemopoietic progenitor cells and in-vitro differentiated erythroblasts, megakaryoblasts, monoblasts and myeloblasts precursors and we analyzed them together with the gene expression profiles of the same populations. We found that for most part of microRNAs specifically up-regulated in one single cell progeny an inverse correlation between microRNAs and down-regulated putative targets expression levels occurs. We chose hsa-mir-299-5p as a model to get further insights into the possible biological relevance of this microRNAs-mRNAs expression integrated analytical approach and we asked if the forced expression of a single lineage-specific microRNA is able to control the cell fate of CD34+ progenitors grown in multilineage culture conditions. Gain and loss of-function experiments established that mir-299-5p regulates hemopoietic progenitors fate modulating reciprocally megakaryocytic-granulocytic versus erythroid-monocytic differentiation and has at least two genuine targets, the transcription factors CTCF and SOX4. CD34+ hematopoietic progenitor cells were transfected with the Amaxa Nucleofector Device, using the Human CD34 Cell Nucleofection Kit, accordingly to the manufacturer’s instructions (Amaxa Biosystem, Cologne, Germany), and 5µg of either the Pre-miR miRNA Precursor Molecule—Negative Control # 1 (NC1) or the hsa-mir-299-5p Pre-miR miRNA Precursor Molecule (299-5p) (Ambion, Austin, TX, USA) and pulsed with the program U-008. The dataset is composed of three independent paired experiment of 299-5p gain of-function (three hsa-299-5p Pre-miR miRNA Precursor Molecule nucleoporated samples and three paired Pre-miR miRNA Precursor Molecule—Negative Control # 1 transfected ones).

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Gene expression signatures in a human bladder cancer cell line SW780 transfected with miR-137 precursor molecule

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