miR‐639 regulates transforming growth factor beta‐induced epithelial–mesenchymal transition in human tongue cancer cells by targeting FOXC1

Epithelial‐to‐mesenchymal transition (EMT) is implicated in embryonic development and various pathological events. Transforming growth factor beta (TGFβ) has been reported to induce EMT in tumor cells, which is a critical step in the process of metastasis leading to cancer spreading and treatment failure. However, the involvement of microRNA during the EMT process in tongue squamous cell carcinoma (TSCC) remains to be determined. To address this question, TSCC cell lines SCC9 and CAL27 were treated with human recombinant TGFβ1 for 48 h. miRNA microarray illustrated that miR‐639 was significantly downregulated in TGFβ‐treated SCC9 cells. Ectopic expression of miR‐639 with miRNA mimics effectively blocked TGFβ‐induced EMT in SCC9 and CAL27 cells, but inhibition of miR‐639 in SCC9 and CAL27 cells with antisense oligonucleotides induced EMT. Computational microRNA target predictions detected a conserved sequence matching to the seed region of miR‐639 in the 3′‐UTR of FOXC1 mRNA. Luciferase reporter assays revealed that miR‐639 targets FOXC1. Ectopic expression of FOXC1 induces EMT in TSCC cells. Silencing FOXC1 expression blocked TGFβ‐induced EMT in SCC9 cells. Clinically, reduced miR‐639 expression was associated with metastasis in TSCC and poor patient survival. The data from the present study suggest that reduced expression of miR‐639 underscores the mechanism of TGFβ‐induced EMT in TSCC by targeting FOXC1 and may serve as therapeutic targets in the process of metastasis. We demonstrated that reduction of miR‐639 is critical for TGF‐b induced EMT and invasiveness of TSCC cells, and ectopic expression of miR‐639 suppresses TGF‐b induced EMT of TSCC cells via silencing FOXC1. More importantly, miR‐639 expression was downregulated in primary TSCCs with lymph node metastasis, and low expression of this miRNA predicts poor patient survival.