The online version of this article (doi:10.1186/1465-9921-15-58) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
MMP, cultured the ASM cells, analyzed the array data, performed the RT-PCRs and drafted the manuscript. ET performed the arrays. PJA performed some of the RT-PCRS. MAL, helped analyze the array data, and helped conceive the study. DSG performed the patient biopsies. IMA & KFA helped with writing the manuscript. All authors read and approved the final manuscript.
The airway smooth muscle (ASM) cell maintains its own proliferative rate and contributes to the inflammatory response in the airways, effects that are inhibited by corticosteroids, used in the treatment of airways diseases.
We determined the differential expression of mRNAs, microRNAs (miRNAs) and long noncoding RNA species (lncRNAs) in primary ASM cells following treatment with a corticosteroid, dexamethasone, and fetal calf serum (FCS).
mRNA, miRNA and lncRNA expression was measured by microarray and quantitative real-time PCR.
A small number of miRNAs (including miR-150, −371-5p, −718, −940, −1181, −1207-5p, −1915, and −3663-3p) were decreased following exposure to dexamethasone and FCS. The mRNA targets of these miRNAs were increased in expression. The changes in mRNA expression were associated with regulation of ASM actin cytoskeleton. We also observed changes in expression of lncRNAs, including natural antisense, pseudogenes, intronic lncRNAs, and intergenic lncRNAs following dexamethasone and FCS. We confirmed the change in expression of three of these, LINC00882, LINC00883, PVT1, and its transcriptional activator, c-MYC. We propose that four of these lincRNAs (RP11-46A10.4, LINC00883, BCYRN1, and LINC00882) act as miRNA ‘sponges’ for 4 miRNAs (miR-150, −371-5p, −940, −1207-5p).
This in-vitro model of primary ASM cell phenotype was associated with the regulation of several ncRNAs. Their identification allows for in-vitro functional experimentation to establish causality with the primary ASM phenotype, and in airway diseases such as asthma and chronic obstructive pulmonary disease (COPD).
Additional file 1: Table S1: Up-regulated mRNAs in healthy ASM cells after stimulation with FCS. Table S2. Down-regulated mRNAs in healthy ASM cells after stimulation with FCS. Table S3. Up-regulated mRNAs in healthy ASM cells after stimulation with Dex + FCS. Table S4. Down-regulated mRNAs in healthy ASM cells after stimulation with Dex + FCS. Table S5. Baseline levels of miRNAs in healthy ASM. Table S6. lncRNAs increased in expression in healthy ASM cells after stimulation with FCS. Table S7. lncRNAs decreased in expression in healthy ASM cells after stimulation with FCS. Table S8. lncRNAs increased in expression in healthy ASM cells after stimulation with Dex + FCS. Table S9. Changes in expression of lncRNAs in healthy ASM cells after stimulation with Dex + FCS. (DOCX 259 KB)
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- Role of non-coding RNAs in maintaining primary airway smooth muscle cells
Mark M Perry
Philip J Austin
Mark A Lindsay
David S Gibeon
Ian M Adcock
Kian Fan Chung
- BioMed Central
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