MiR-9-5p promotes cell growth and metastasis in non-small cell lung cancer through the repression of TGFBR2

Background: Increasing evidence indicates that the dysregulation of microRNAs (miRNAs) play critical roles tumor progression and metastasis, but very few papers had reported the function of miR-9-5p in lung cancer, especially in NSCLCs. Methods: In this study, we investigated the role of miR-9-5p in non-small cell lung cancers (NSCLCs). MiR-9-5p level were analyzed in 62 clinical NSCLC lung tissue samples and adjacent normal lung tissues by RT-PCR. The target of miR-9-5p was predicted by TargetScan and luciferase reporter assay was used to verify the binding site of miR-9-5p on TGFBR2 mRNA. MTT assay, wound healing assay and invasion assay were performed in both miR-5p inhibitor transfected A549 and miR-5p mimic transfected SK-MES-1 cells. To further investigate whether TGFBR2 is the major target of miR-9-5p, we used TGFBR2 siRNA to transfect A549 and SK-MES-1 cells with miR9-5p inhibitor or miR-9-5p mimic transfection. Western blot were then used to analyze TGFBR2, p-smad2 and psmad3 protein expressions after transfection. Results: Results indicated that NSCLC patients' tissues had a significantly higher expression of miR-9-5p compared to adjacent normal lung tissues. MiR-9-5p mimic transfection promoted proliferation, metastasis and invasion abilities in both A549 and SK-MES-1 cells. Conversely, miR-9-5p inhibitor transfection showed the decreased abilities of these cells. Luciferase reporter assay indicated that TGFBR2 is a direct target of miR-9-5p and the up-regulation of TGFBR2 suppressed cell proliferation, metastasis and invasion. The knock down of TGFBR2 abrogated the effect of miR-9-5p in down-regulating p-smad2 and p-smad3 expressions, which indicated that TGFBR2 is the major target of miR-9-5p in NSCLC cells. Conclusions: Our finding indicated that miR-9-5p promotes the proliferation, metastasis and invasion of NSCLC cells by down-regulating TGFBR2 expression.