摘要
目的探索造血促进因子白细胞介素（interleukin，IL）-11与甲状腺未分化癌（anaplastic thyroid carcinoma，ATC）进展间的关系。方法采用ELISA和Real-time PCR分别检测甲状腺癌细胞系上清IL-11蛋白水平和mRNA水平；构建稳定敲低IL-11的ATC细胞株，采用MTY、Transwell、划痕实验及Western blot方法分析敲低IL-11或外源性重组人巨细胞介素（recombinant human interleukin，rhIL）-11对ATC细胞株增殖、侵袭和迁移能力的影响及机制。使用SPSS 19．0软件统计分析数据，各组之间均值的比较采用Student’t检验。结果ELISA方法检测发现3个干扰载体组细胞中IL-11蛋白表达量较空载体组显著低（21．55±1．69、16．18±0．85、26．37±2．00比54．54±3．99，P值均〈0．05）；平板克隆实验发现IL-11敲低组细胞与对照组sw579细胞形成克隆数目差异无统计学意义（P〉0．05），同时MTr实验发现，敲低IL—11后其细胞增殖速度与对照组sw579细胞差异无统计学意义（P〉0．05）；Transwell侵袭和迁移实验发现外源性rhIL-11（0～100ns／m1）处理ATC细胞后，穿过小室的细胞随着rhIL-11浓度的升高而增多。结论IL-11通过介导ATC细胞上皮间质转化促进其侵袭转移能力，IL-11可作为ATC分子靶向治疗的一个潜在靶点。
Objective To explore the effect of hematopoietic cytokines IL-11 on invasion and metastasis abilities of anaplastic thyroid caeinoma（ATC） cells. Methods Real-time PCR was performed for examining the IL-11 mRNA expression in thyroid carcinoma cell lines, and IL-11 protein expression in the supernament of thyroid carcinoma cell lines was detected by ELISA. Molecular cloning was employed to construct IL-11 stable knockdown cell line; MTT assay was used to analyze the effect of IL-11 on the proliferation of ATC ceils; Transwell and wound healing assays were employed to analyze the abilities of migration and invasion in ATC cells. Western blotting was used to detect the relative pathway proteins. SPSS statistical package 19.0 was used to analyze the date, and Student＇s t test was used for multiple comparisons. Results The protein level of IL-11 were significantly lower in knock-down cell lines than that in negative control cell lines（21.55 ± 1.69, 16.18 ± 0.85, 26.37 ± 2.00 vs 54.54 _＋ 3.99, all P 〈 0.05 ）. Colony formation assays reveal that colony number between knock-down cells and negative control cells has no significance（ P 〉 0. 05 ）. Meanwhile, MTT assays show that there is no significance between knock-down cell lines and negative control cell line （P 〉 0.05 ）. However, Transwell invasion and migration assays show that number of migrated cells is increased when ATC cells were treated with rhIL-11 （0-100 ng/ml）at increasing concentrations. Conclusion IL-11 improves the migratory and invasive abilities of ATC cells via inducing EMT of ATC ceils, and it can be used as a potential target for ATC molecular targeted therapy.
作者
蒋玉娥
钟兆铭
陈雪
王伟
罗娟章
王士琪
储红映
孙传政
Clinical Laboratory, the Third Affiliated Hospital of Kunming Medical University, Kunming 650118, China(Jiang YE ） ; Department of Head and Neck Surgery, the Third Affiliated Hospital of Kunming Medical University, Kunming 650118, China （Zhong ZM, Chen X, Wang W, Luo JZ, Wang SQ, Chu HY, Sun CZ ） ; Department of Oncology, the First Affiliated Hospital of Kunming Medical University, Kunming 650118, China（ Zhong ZM)
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2018年第4期286-291,共6页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金
国家自然科学基金（81560470、81773127）
云南省中青年学术技术带头人后备人才专项经费（2015HB086）
关键词
甲状腺肿瘤
白细胞介素-11
上皮间质转化
侵袭与转移
Thyroid neoplasms
Interleukin-ll
Epithelial mesenchymal transition
Invasion and metastasis
