MiR-200a inhibits migration of NSCLC cell line A549 by targeting Sulfatase 2 / 中华胸心血管外科杂志

ABSTRACT

Objective To investigate the effect of miR-200a on the migratory ability of NSCLC cells and to explore its possible mechanism.Methods Real-time PCR was performed to analyze the miR-200a expression in NSCLC cell lines A549 and SK-MES-1, and human normal lung bronchial epithelial cell line 16HBE.Hsa-miR-200a mimics, NC mimics, hsa-miR-200a inhibitor and NC inhibitor were transfected into A549 cells using Lipofectamine 2000.Migration of A549 cells was detected by Transwell migration assay.The potential target genes of miR-200a were predicted by bioinformatics software and then verified by dual luciferase reporter gene assay and Western blot.Results MiR-200a was significantly down-regulated in A549 and SK-MES-1 cells(P＜0.05).Exogenous over-expression of miR-200a mimics significantly inhibited migratory a-bility of A549 cells, while over-expression of miR-200a inhibitor generated the opposite effect(P＜0.01).Dual luciferase re-porter assay indicated that miR-200a could directly affect the 3'-UTR of Sulf2 gene to inhibit luciferase activity.Western blot revealed that miR-200a expression could significantly reduce Sulf2 protein expression level in A549 cells.Ectopic expression of Sulf2 protein in miR-200a-overexpressing A549 cells overrode the migration inhibition effect of miR-200a, suggesting that targe-ting Sulf2 represents an important mechanism of the anti-tumour activity of miR-200a in lung cancer.Conclusion MiR-200a inhibits migration of lung cancer cells by targeting Sulf2.