miR-221 participates in the airway epithelial cells injury in asthma via targeting SIRT1.

AIM OF THE STUDY

To investigate the role of microRNA-221 (miR-221) in the airway epithelial cell injury in asthma and delineate the underlying mechanism that may involve with SIRT1.

MATERIALS AND METHOD

Bronchial epithelial cells from asthma patients and healthy controls were obtained by bronchoscopic brushing. The miR-221 and SIRT1 mRNA level in collected cells were detected by qRT-CPR. BEAS2B cell lines were cultured in vitro. In order to up-regulate miR-221 and SIRT1, miR-221 mimic and pcDNA3.1-SIRT1 vector was transfected into BEAS2B cells, respectively. The expression changes of miR-221 and SIRT1 after transfection was observed by qRT-PCR and Western blot. The target relationship between miR-221 and SIRT1 was confirmed using dual-luciferase reporter assay.The cell viability changes after transfection was measured using cellTiter-blue reagent. The apoptosis rate was detected by flow cytometry.

RESULT

Compared with healthy controls, miR-221 expression significantly increased in bronchial epithelial cells from patients subjects. In contrast, the level of SIRT1 mRNA reduced in the bronchial epithelial cell from asthma patients. In vitro, up-regulation of miR-221 could inhibit the expression of SIRT1 both at mRNA and protein level in BEAS2B cells. A negative correlation between miR-221 and SIRT1 mRNA in samples from patients was confirmed and dual-luciferase reporter assay showed that miR-221 directly binds to the 3'UTR of SIRT1 mRNA. Overexpression of miR-221 or SIRT1 knockdown could inhibit proliferation but induce apoptosis in BEAS2B cells. Moreover, up-regulation of SIRT1 could antagonize miR-221's inhibitory effect.

CONCLUSION

miR-221 may participate in the airway epithelial cells injury in asthma via targeting SIRT1.