DIO1抑制喉鳞状细胞癌侵袭转移的机制研究  

Study on the mechanism of DIO1 inhibiting invasion and metastasis in laryngeal squamous cell carcinoma

作　　者：任楠 刘金婧 徐海铭 赵凯 刘明波[1] REN Nan;LIU Jinjing;XU Haiming;ZHAO Kai;LIU Mingbo(Department of Otolaryngology Head and Neck Surgery,Chinese People’s Liberation Army General Hospital,Beijing 100853,China;Department of Otolaryngology Head and Neck Surgery,Hainan Hospital of Chinese People’s Liberation Army General Hospital,Hainan Province,Sanya 572013,China)

机构地区：[1]解放军总医院耳鼻咽喉头颈外科医学部,北京100853 [2]解放军总医院海南医院耳鼻咽喉头颈外科,海南三亚572013

出　　处：《中国医药导报》2023年第18期25-29,共5页China Medical Herald

基　　金：中央引导地方科技发展专项资金项目(ZY2021HN03);海南省科技计划项目(LCYX202105)。

摘　　要：目的研究碘甲原氨酸脱碘酶Ⅰ(DIO1)在喉鳞状细胞癌(LSCC)侵袭转移中的调控作用。方法将TU686细胞分为对照组(正常TU686细胞)和DIO1过表达组(转染DIO1),并根据过表达量继续分为DIO1-1μg组、DIO1-2μg组。RT-qPCR检测DIO1基因表达,CCK-8实验检测细胞增殖能力,PI/DAPI染色检测细胞凋亡情况,划痕实验及Transwell实验检测细胞迁移及侵袭能力,基因探针富集分析相关基因集的富集情况,Western blot检测DIO1、E-cadherin、N-cadherin、Vimentin、p-S6K1、p-4EBP1、p-Akt蛋白表达水平。结果DIO1过表达组DIO1 mRNA、蛋白水平高于对照组(P<0.01)。24、48、72 h,DIO1过表达组光密度值低于对照组(P<0.01)。DIO1过表达组PI染色阳性比例高于对照组(P<0.01)。DIO1过表达组相对迁移距离短于对照组(P<0.01)。DIO1过表达组迁移及侵袭能力低于对照组(P<0.01)。DIO1表达与上皮间质转化、mTORC1信号通路相关基因集的表达呈负相关(NES<0)。DIO1-1μg组、DIO1-2μg组E-cadherin高于对照组,N-cadherin、Vimentin、p-S6K1和p-4EBP1低于对照组(P<0.01)。DIO1-1μg组、DIO1-2μg组与对照组p-Akt比较,差异无统计学意义(P>0.05)。结论DIO1可作为抑癌基因,在LSCC侵袭转移中起关键作用。Objective To study the regulatory effect of deiodinase iodothyronine typeⅠ(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 DIO1-1μg group and DIO1-2μg 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 DIO1-1μg group and DIO1-2μg 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 DIO1-1μg group,DIO1-2μg group(P>0.05).Conclusion DIO1 can be a tumor suppressor gene and play a key role in the invasion and metastasis of LSCC.

关 键 词：喉鳞状细胞癌 碘甲原氨酸脱碘酶Ⅰ 侵袭与转移 

分 类 号：R739.65[医药卫生—肿瘤]