Unknown,Transcriptomics,Genomics,Proteomics

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MiRNA expression in cytokine-stimulated airway smooth muscle cells

ABSTRACT: We used microarrays to determine if miRNA expression in human airway smooth muscle cells is altered by a pro-inflammatory stimulus. A majority of the miRNAs on the array exhibited very low signal intensity. In ASM cells exposed to IL-1Î², TNFÎ±, and IFNÎ³, none of the miRNA expressed were significantly up-regulated with cytokine treatment. We did observe 11 miRNA down-regulated > 2-fold in both cultures with cytokine treatment. These miRNA include described human miRNA miR-23a, -23b, -25, -188, -320, -363, -489, as well as mouse and rat miRNA homologous to miR-140*, mouse miR-329 and a novel miRNA, abi-13268. miRNA arrays for cytokine-stimulated ASM cells were completed in duplicate from two different cultures to identify candidate miRNA for further study. Cultures were growth arrested for 48 hours and treated with 10 ng/ml IL-1Î², TNFÎ±, and IFNÎ³ for 24 hrs. A comparison of miRNA expression under cytokine-stimulated and non-treated conditions from hybridized miRNA arrays was calculated as described.

ORGANISM(S): Homo sapiens

SUBMITTER: Cherie Singer

PROVIDER: E-GEOD-16587 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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MicroRNA expression in human airway smooth muscle cells: role of miR-25 in regulation of airway smooth muscle phenotype.

Kuhn Andrew R AR Schlauch Karen K Lao Ronna R Halayko Andrew J AJ Gerthoffer William T WT Singer Cherie A CA

American journal of respiratory cell and molecular biology 20090618 4

Defining mechanisms by which differentiated, contractile smooth muscle cells become proliferative and secretory in response to mechanical and environmental stress is crucial for determining the contribution of airway smooth muscle (ASM) to inflammatory responses that result in airway disease. Regulation by microRNAs (miRNAs) has emerged as an important post-transcriptional mechanism regulating gene expression that may modulate ASM phenotype, but little is known about the expression and functions ...[more]

PMID: 19541842

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Project description:miRNA expression was compared in skin from control newborn mice and littermate mice ectopically expressing the potent secreted WNT inhibitor Dickkopf 1 (DKK1) in the epidermis. DKK1 completely suppresses hair follicle development. Multiple miRNAs were identified that reproducibly produced signals above background in both types of skin sample. Several miRNAs were identified for which hybridization signals were on average more than 2.5 fold higher in control samples than in Dkk1-expressing samples, suggesting these may be upregulated in hair follicles, and/or are direct or indirect targets of WNT inhibition in the skin. Keywords: miRNA expression array, transgenic mouse, skin, WNT Low molecular weight RNA was isolated using the mirVana RNA extraction kit (Ambion) from full thickness dorsal skin of three K5-rtTA; tetO-Dkk1 newborn mice and three control littermates following doxycycline treatment from E0.5 to induce expression of Dkk1 in double transgenic epidermis [1]. Control and Dkk1âexpressing transgenic samples were tagged for labeling with Cy3 and Cy5 dyes respectively using the Ncode miRNA Labeling System (Invitrogen), mixed by equal mass, and co-hybridized to miRNA arrays that have been described previously [2]. Background-corrected median signals for pixels from each probe spot in both the Cy3 and Cy5 channels were used for analysis. Cy3 (control samples C2, C4, C5) was detected at 532nm and Cy5 (Dkk1âexpressing transgenic samples TG1, TG2, TG4) at 635 nm. References: [1] Chu, E.Y., Hens, J., Andl, T., Kairo, A., Yamaguchi, T.P., Brisken, C., Glick, A., Wysolmerski, J.J., and Millar, S.E. (2004). Canonical WNT signaling promotes mammary placode development and is essential for initiation of mammary gland morphogenesis. Development 131, 4819-4829. [2] Nelson, P.T., Baldwin, D.A., Scearce, L.M., Oberholtzer, J.C., Tobias, J.W., and Mourelatos, Z. (2004). Microarray-based, high-throughput gene expression profiling of microRNAs. Nat Methods 1, 155-161.

Dataset Information

MiRNA expression in cytokine-stimulated airway smooth muscle cells

Publications

MicroRNA expression in human airway smooth muscle cells: role of miR-25 in regulation of airway smooth muscle phenotype.

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