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Published on: May 27, 2015
MicroRNA-143 downregulates interleukin-13 receptor alpha1 in human mast cells
Shaoqing Yu1, Ruxin Zhang, Chunshen Zhu
1Department of Otolaryngology, Huadong Hospital, Fudan University, Shanghai 200040, China. yu_shaoqing@163.com
Abstract:
MicroRNA-143 (miR-143) was found to be downregulated in allergic rhinitis, and bioinformatics analysis predicted that IL-13Rα1 was a target gene of miR-143. To understand the molecular mechanisms of miR-143 involved in the pathogenesis of allergic inflammation, recombinant miR-143 plasmid vectors were constructed, and human mast cell-1(HMC-1) cells which play a central role in the allergic response were used for study. The plasmids were transfected into HMC-1 cells using a lentiviral vector. Expression of IL-13Rα1 mRNA was then detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western Blotting. The miR-143 lentiviral vector was successfully stably transfected in HMC-1 cells for target gene expression. Compared to the control, the target gene IL-13Rα1 was less expressed in HMC-1 transfected with miR-143 as determined by RT-PCR and Western Blotting (p < 0.05); this difference in expression was statistically significant and the inhibition efficiency was 71%. It indicates that miR-143 directly targets IL-13Rα1 and suppresses IL-13Rα1 expression in HMC-1 cells. Therefore, miR-143 may be associated with allergic reaction in human mast cells.
Insights
MicroRNA-143 (miR-143) suppresses IL-13Rα1 expression in human mast cells, suggesting a role in allergic inflammation. This finding offers potential therapeutic targets for allergic rhinitis.
Area of Science:
- Molecular Biology
- Immunology
- Genetics
Background:
- Allergic rhinitis involves complex immune responses.
- MicroRNA-143 (miR-143) is downregulated in allergic rhinitis.
- Interleukin-13 receptor subunit alpha 1 (IL-13Rα1) is a predicted target of miR-143.
Purpose of the Study:
- To investigate the molecular mechanisms of miR-143 in allergic inflammation.
- To confirm if miR-143 directly targets and suppresses IL-13Rα1 expression.
- To understand the role of miR-143 in human mast cells (HMC-1) during allergic reactions.
Main Methods:
- Construction of recombinant miR-143 plasmid vectors using lentiviral vectors.
- Stable transfection of miR-143 lentiviral vectors into HMC-1 cells.
- Quantification of IL-13Rα1 mRNA and protein expression using RT-PCR and Western Blotting.
Main Results:
- Successful stable transfection of miR-143 lentiviral vectors into HMC-1 cells.
- Significantly reduced expression of IL-13Rα1 mRNA and protein in miR-143 transfected HMC-1 cells compared to controls (p < 0.05).
- An inhibition efficiency of 71% for IL-13Rα1 expression by miR-143 was observed.
Conclusions:
- miR-143 directly targets IL-13Rα1 and suppresses its expression in HMC-1 cells.
- miR-143 plays a role in regulating allergic inflammation through human mast cells.
- These findings suggest miR-143 as a potential therapeutic target for allergic diseases.
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