炎症因子通过核因子κB通路促进口腔鳞状细胞癌转移的体外研究  

Inflammatory factors promote oral squamous cell carcinoma cells metastasis, via nuclear factor kappa B signal pathway in vitro

作　　者：严明[1] 何地[1] 张萍[1] 周晓健[1] 陈万涛[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔医学院口腔颌面外科上海市口腔医学研究所上海市口腔医学重点实验室,200011

出　　处：《中华口腔医学杂志》2010年第3期146-151,共6页Chinese Journal of Stomatology

基　　金：上海市科学技术委员会科研计划（08JC1414400、09431902200、10XD1402500）;上海市重点学科研究项目（S30206）

摘　　要：目的 探讨炎症因子与核因子κB（nuclear factor kappa B,NF-κB）信号转导通路在口腔鳞状细胞癌转移中的意义.方法 应用口腔鳞状细胞癌低转移细胞系Tca8113和高转移细胞系Tb为研究对象,通过蛋白质印迹法和荧光素酶报告基因法检测口腔鳞状细胞癌细胞系中NF-κB通路活性.用NF-κB抑制因子α（inhibitor of kappa B alpha,IκBa）抑制质粒第32、36位丝氨酸磷酸化位点被丙氨酸替代的质粒（pBabe-SR-IκBa）和NF-κB通路抑制剂吡咯烷二硫代氨基甲酸盐（pyrolidinedithiecar bamate,PDTC）抑制信号转导通路活性,并用侵袭小室实验（TransweH）检测口腔鳞状细胞癌高转移系Tb细胞侵袭能力的变化.此外,通过酶联免疫吸附法检测pBabe-SR-IκBα和PDTC抑制信号转导通路后,肿瘤坏死因子α（tumor necrosis factor-alpha,TNF-α）、白介素（IL）-1a、IL-6、IL-8和粒-巨噬细胞集落刺激因子（granulocyte-macrophage colony stimulating factor,GM-CSF）等炎症因子的分泌.结果 蛋白质印迹法检测显示：高转移Tb细胞中磷酸化IκBα和磷酸化p65的表达量明显高于低转移细胞系Tea8113,分别为Tea8113细胞的3.19倍和6.81倍.荧光素酶（luciferase）报告基因结果显示,Tb细胞NF-κB的启动子活性为Tca8113的2.12倍（P〈0.01）,并对TNF-α更为敏感.转染pBabe-SR-IκBα和应用PDTC抑制NF-κB通路后,对Tb细胞的体外侵袭能力抑制率分别为21.9%和69.3%.此外,酶联免疫吸附试验法显示通路抑制后,TNF-α、IL-1α、IL-6、IL-8和GM-CSF等炎症因子的分泌也明显降低.结论 TNF-α、IL-1α、IL-6、IL-8和GM-CSF等炎症因子可能通过NF-κB通路促进口腔鳞状细胞癌细胞转移.Objective To investigate the mles of inflammatory factors and nuclear factor kappa B （NF-κB）signal pathway in metastasis of oral squamous eell carcinoma.Methods The oral squamous cell carcinoma cell lines with highly metastasis potential （Tb） and lower metastasis potential （Tca8113） were used in this study.The levels of NF-κB activity in oral squamous cell carcinoma cell lines were determined by Westem blotting and luciferase reporter assay.pBabe-IκBα-SR expression vector or NF-κB inhibitor pyrolidinedithiocarbamate （PDTC） was used to inhibit NF-κB,and cell migration was examined by transwell assay.The secretion of tumor necrosis factor.alpha （TNF-α）,IL-1α,IL-6,IL-8 and GM-CSF proinflammatory cytokines was determined by ELISA when Tb cells were transfected with pBabe-SR-IκBα or treated with PDTC.Results Western blotting showed that the levels of phosphorIκBα and phosphor-p65 were highiy expressed in Tb ceus.Tb cells had high level of constitutive NF-κB activity and were more sensitive tu TNF-α.The migration of highly metastatic Tb cells,either transfected with dominant-negative mutant inhibitor pBαbe-SR-IκBα or treated with PDTC,Waft;suppressed when determined by transwell assay.The secretion of proinflammatory cytokines,including TNF-α,IL-1α,IL-6,IL-8 and granulocytemacrophage colony stimulating factor （GM-CSF）,was inhibited by pBαbe-SR-IκBα transfection or PDTC treatment.Conclusion The inflammatory factors such as TNF-α could promote oral squamous cell carcinoma cell metastasis via NF-κB signal pathway.

关 键 词：核因子ΚB 炎症趋化因子类 癌 鳞状细胞 肿瘤转移 

分 类 号：R739.8[医药卫生—肿瘤]