Hypoxia Induces Epithelial-Mesenchymal Transition in Follicular Thyroid Cancer: Involvement of Regulation of Twist by Hypoxia Inducible Factor-1 alpha

Abstract

Purpose: Although follicular thyroid cancer (FTC) has a relatively fair prognosis, distant metastasis sometimes results in poor prognosis and survival. There is little understanding of the mechanisms contributing to the aggressiveness potential of thyroid cancer. We showed that hypoxia inducible factor-1 alpha (HIF-1 alpha) induced aggressiveness in FTC cells and identified the underlying mechanism of the HIF-1 alpha-induced invasive characteristics. Materials and Methods: Cells were cultured under controlled hypoxic environments (1% O-2) or normoxic conditions. The effect of hypoxia on HIF-1 alpha, and epithelial-to-mesenchymal transition (EMT) related markers were evaluated by quantitative real-time PCR, Western blot analysis and immunocytochemistry. Invasion and wound healing assay were conducted to identify functional character of EMT. The involvement of HIF-1 alpha and Twist in EMT were studied using gene overexpression or silencing. After orthotopic nude mouse model was established using the cells transfected with lentiviral shHIF-1 alpha, tissue analysis was done. Results: Hypoxia induces HIF-1 alpha expression and EMT, including typical morphologic changes, cadherin shift, and increased vimentin expression. We showed that overexpression of HIF-1 alpha via transfection resulted in the aforementioned changes without hypoxia, and repression of HIF-1 alpha with RNA interference suppressed hypoxia-induced HIF-1 alpha and EMT. Furthermore, we also observed that Twist expression was regulated by HIF-1 alpha. These were confirmed in the orthotopic FTC model. Conclusion: Hypoxia induced HIF-1 alpha, which in turn induced EMT, resulting in the increased capacity for invasion and migration of cells via regulation of the Twist signal pathway in FTC cells. These findings provide insight into a possible therapeutic strategy to prevent invasive and metastatic FTC.