Xiaowen Zhang1,2, Nan Wu3, Jin Wang4. 1. Medical Research Center, Shengjing Hospital of China Medical University, #36 Sanhao Street, Heping District, Shenyang, 110004, China. zhangxiaowen@cmu.edu.cn. 2. Key Laboratory of Research and Application of Animal Model for Environmental and Metabolic Diseases, Shenyang, China. zhangxiaowen@cmu.edu.cn. 3. The Central Laboratory of the First Affiliated Hospital of China Medical University, Shenyang, China. 4. The ENT Department, Shengjing Hospital of China Medical University, Shenyang, China.
Abstract
PURPOSE: A new circular RNA (hsa-circ_0058106) has been found to be upregulated in laryngeal cancer, but its function remains unknown. Here, we explored its role in the metastasis of laryngeal cancer. METHODS: The level of hsa-circ_0058106 in laryngeal cancer was detected by qRT-PCR. The effect of hsa-circ_0058106 silencing on the metastasis of laryngeal cancer was assessed using scratch wound healing and transwell assays, as well as nude mouse lung metastasis models. Fluorescence in situ hybridization was employed to analyze the cellular localization of hsa-circ_0058106. A luciferase activity assay was used to assess binding between hsa-circ_0058106 and miR-153. Interaction between hsa-circ_0058106 and Twist1 was confirmed using RNA pull-down and RNA immunoprecipitation assays. RESULTS: A high level of hsa-circ_0058106 was found to be associated with lymph node metastasis and advanced clinical stages. Stable knockdown of hsa-circ_0058106 inhibited the migration and invasion of laryngeal cancer cells in vitro and in vivo. Moreover, we found that downregulation of hsa-circ_0058106 suppressed epithelial-mesenchymal transition (EMT), which was underscored by the observation that hsa-circ_0058106 silencing led to decreases in the expression of N-cadherin and Vimentin, and an increase in the expression of E-cadherin. Mechanistically, we found that hsa-circ_0058106 can specifically bind to miR-153 and regulate Snail1 expression by acting as a miR-153 sponge. In addition, we found that knockdown of hsa-circ_0058106 blocked the nuclear translocation of Twist1. CONCLUSIONS: Our results indicate that hsa-circ_0058106 induces EMT and metastasis in laryngeal cancer by sponging miR-153 and inducing Twist1 nuclear translocation. These observations provide new insights into the regulatory effects of circRNAs in laryngeal cancer metastasis.
PURPOSE: A new circular RNA (hsa-circ_0058106) has been found to be upregulated in laryngeal cancer, but its function remains unknown. Here, we explored its role in the metastasis of laryngeal cancer. METHODS: The level of hsa-circ_0058106 in laryngeal cancer was detected by qRT-PCR. The effect of hsa-circ_0058106 silencing on the metastasis of laryngeal cancer was assessed using scratch wound healing and transwell assays, as well as nude mouse lung metastasis models. Fluorescence in situ hybridization was employed to analyze the cellular localization of hsa-circ_0058106. A luciferase activity assay was used to assess binding between hsa-circ_0058106 and miR-153. Interaction between hsa-circ_0058106 and Twist1 was confirmed using RNA pull-down and RNA immunoprecipitation assays. RESULTS: A high level of hsa-circ_0058106 was found to be associated with lymph node metastasis and advanced clinical stages. Stable knockdown of hsa-circ_0058106 inhibited the migration and invasion of laryngeal cancer cells in vitro and in vivo. Moreover, we found that downregulation of hsa-circ_0058106 suppressed epithelial-mesenchymal transition (EMT), which was underscored by the observation that hsa-circ_0058106 silencing led to decreases in the expression of N-cadherin and Vimentin, and an increase in the expression of E-cadherin. Mechanistically, we found that hsa-circ_0058106 can specifically bind to miR-153 and regulate Snail1 expression by acting as a miR-153 sponge. In addition, we found that knockdown of hsa-circ_0058106 blocked the nuclear translocation of Twist1. CONCLUSIONS: Our results indicate that hsa-circ_0058106 induces EMT and metastasis in laryngeal cancer by sponging miR-153 and inducing Twist1 nuclear translocation. These observations provide new insights into the regulatory effects of circRNAs in laryngeal cancer metastasis.