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| Study on the mechanism of DIO1 inhibiting invasion and metastasis in laryngeal squamous cell carcinoma |
| REN Nan1 LIU Jinjing1 XU Haiming1 ZHAO Kai2 LIU Mingbo1 |
1.Department of Otolaryngology Head and Neck Surgery, Chinese People’s Liberation Army General Hospital, Beijing 100853, China;
2.Department of Otolaryngology Head and Neck Surgery, Hainan Hospital of Chinese People’s Liberation Army General Hospital, Hainan Province, Sanya 572013, China
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Abstract Objective To study the regulatory effect of deiodinase iodothyronine type Ⅰ (DIO1) on invasion and metastasis in laryngeal squamous cell carcinoma (LSCC). Methods The TU686 cells were divided into control group (normal TU686 cells) and DIO1 overexpression group (transfection of DIO1), and DIO1-1 μg group and DIO1-2 μg group were further divided according to overexpression level. DIO1 gene expression was detected by RT-qPCR, cell proliferation was detected by CCK-8 assay, cell apoptosis was detected by PI/DAPI staining, cell migration and invasion ability were detected by scratch assay and Transwell assay, the enrichment of related gene sets was analyzed by gene probe enrichment analysis, and the protein expression levels of DIO1, E-cadherin, N-cadherin, Vimentin, p-S6K1, p-4EBP1, and p-Akt were detected by Western blot. Results DIO1 mRNA and protein level of DIO1 overexpression group were higher than those of control group (P＜0.01). At 24, 48, 72 h, optical density values of DIO1 overexpression group were lower than those of control group (P＜0.01). The positive percentage of PI staining in DIO1 overexpression group was higher than that in control group (P＜0.01). The relative migration distance of DIO1 overexpression group was shorter than control group (P＜0.01).Migration and invasion ability of DIO1 overexpression group was lower than control group (P＜0.01). DIO1 expression was negatively correlated with the expression of epithelial-mesenchymal transition and mTORC1 signaling pathway genes(NES＜0). E-cadherinin DIO1-1 μg group and DIO1-2 μg group were higher than those in control group, while N-cadherin and Vimentin, p-S6K1， and p-4EBP1 were lower than those in control group (P＜0.01). There were no significant differences in p-Akt between control group and DIO1-1 μg group, DIO1-2 μg group (P＞0.05). Conclusion DIO1 can be a tumor suppressor gene and play a key role in the invasion and metastasis of LSCC.
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