甲状腺癌患者外周血及肿瘤组织中IL-8的表达及其作用的初步研究  

A preliminary study on expression and role of IL-18 in peripheral blood and tumor tissues of patients with thyroid cancer

作　　者：沈小波[1] 任汉强[1] 陈伟[1] 龚传明 SHEN Xiao-Bo;REN Han-Qiang;CHEN Wei;GONG Chuan-Ming(Endocrine Department,Affiliated Hospital ofJianghan University,Wuhan 430015,China)

机构地区：[1]江汉大学附属医院内分泌科,武汉430015 [2]中国人民解放军161医院,武汉430015

出　　处：《中国免疫学杂志》2018年第8期1237-1241,1250,共6页Chinese Journal of Immunology

摘　　要：目的:通过分析甲状腺癌患者IL-18的表达探讨IL-18对甲状腺癌的作用。方法:本研究共纳入2015年6月~2017年1月我院收治的123例甲状腺癌患者作为研究对象。另选取20例同期在我院进行体检的健康者作为对照。采用ELISA法(酶联免疫吸附法)检测甲状腺癌患者和健康者的血清IL-18水平。采用Western blot和免疫组化分析甲状腺癌患者癌组织及癌旁组织中IL-18、IFN-γ及p-STAT1的表达。建立小鼠模型,采用TRK-T1转基因小鼠诱导甲状腺癌,构建TRK-T1单转基因与TRK-T1/IL-18双转基因小鼠模型,将单转基因与双转基因后代小鼠进行比较。记录两组小鼠的甲状腺癌发生率,绘制小鼠的生存曲线。采用Western blot和免疫组化分析各组小鼠甲状腺癌组织中IL-18、IFN-γ及p-STAT1的表达水平。流式细胞术检测各组小鼠甲状腺癌组织中CD4+T细胞、CD8+T细胞与CD56+CD16+NK细胞数量。分离正常C57BL/6J小鼠的外周血单核细胞(PBMC),分别转染IFN-γ及对照siRNA,使用IL-18处理后检测NK细胞的活性水平。结果:ELISA结果显示,甲状腺癌患者的血清IL-18水平显著高于健康者(P<0.05)。Western blot和免疫组化分析显示甲状腺癌患者癌组织中IL-18、IFN-γ及p-STAT1的表达水平均显著高于癌旁组织(P<0.05)。TRK-T1单转基因与TRK-T1/IL-18双转基因小鼠的甲状腺癌发病率分别为27.3%和7.5%,两者相比差异具有统计学意义(P<0.05)。TRK-T1/IL-18双转基因小鼠的生存率显著优于TRK-T1单转基因小鼠(P<0.05)。Western blot和免疫组化分析显示TRK-T1/IL-18双转基因小鼠甲状腺癌组织中IL-18、IFN-γ及p-STAT1的表达水平均显著高于TRK-T1单转基因小鼠(P<0.05)。流式分析显示TRK-T1/IL-18双转基因小鼠甲状腺癌组织中CD4+T细胞、CD8+T细胞与CD56+CD16+NK细胞数量均显著高于TRK-T1单转基因小鼠(P<0.05)。IL-18处理能显著增加正常PBMC中NK细胞的活性(P<0.05),而不影响转染IFN-γsiRNA的PBMC中NK细胞的�Objective： To explore the role of IL-18 in thyroid cancer by investigating the expression of IL-18 in peripheral blood and tumor tissues of patients with thyroid cancer. Methods： A total of 123 cases of patients with thyroid cancer underwent surgical treatment in our hospital from June 2015 to January 2017 were recruited as study object. Another 20 cases of healthy people who underwent physical examination in the physical examination center of our hospital were selected as control. Serum levels of IL-18 in patients with thyroid cancer and healthy people were determined using a commercially available sandwich ELISA（ Enzyme-Linked Immuno Sorbent Assay）. Western blot and immunohistochemical staining（ IHC） were used to detect the expression level of IL-18,IFN-γ and p-STAT1 in tumor and paracarcinoma tissue. For mice model,TRK-T1 transgenic mice were used to induce thyroid cancer,construct TRK-T1 single transgenic and TRK-T1/IL-18 double transgenic mice,the double transgenic mice were compared with the single transgenic mice. The incidence of thyroid cancer in different group were recorded and the survival curves were drawn. Western blot and IHC were used to detect the expression level of IL-18,IFN-γ and p-STAT1 in tumor tissue of mice in different groups. Flow cytometry was used to determine the number of CD4＋T,CD8＋T and CD56＋CD16＋NK cells in tumor tissue. The peripheral blood mononuclear cell（ PBMC） of C57 BL/6 J mice were separated,IFN-γ and normal control siRNA were transfected then,the activity of NK cells were determined after treatment of IL-18. Results： The results of ELISA showed that the serum level of IL-18 in patients with thyroid cancer were significant higher than in healthy people（ P〈0. 05）. The results of Western blot and IHC showed that the expression level of IL-18,IFN-γ and pSTAT1 in tumor were significant higher than in paracarcinoma tissue（ P 〈 0. 05）. The incidence of thyroid cancer of TRK-T1 single transgenic and TRK-T1/IL-18 double transgenic mice were

关 键 词：IL-18 IFN-Γ STAT1 甲状腺癌 

分 类 号：R736[医药卫生—肿瘤]