MUC1蛋白在侵袭性垂体腺瘤表达及其诱导抗肿瘤免疫反应的研究  

Expression of MUC1 protein in invasive pituitary adenomas and its role in anti-tumor immune response

作　　者：马立新[1,3] 刘志慧[2] 隋德华[1] 徐文虎[1] 王波[1] 邢海龙[1] 李勐[1] 李泽福[1] 

机构地区：[1]滨州医学院附属医院神经外科,滨州256603 [2]滨州医学院附属医院妇产科,滨州256603 [3]山东大学齐鲁医院神经外科,济南250012

出　　处：《中华神经医学杂志》2015年第12期1250-1255,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81171119）

摘　　要：目的探讨MUC1蛋白在侵袭性和非侵袭性垂体腺瘤中的表达及其诱导的抗肿瘤免疫反应。方法收集滨州医学院附属医院及山东大学齐鲁医院神经外科自2009年3月至2014年12月手术切除的人垂体腺瘤标本86例，其中侵袭性垂体腺瘤42例，非侵袭性垂体腺瘤44例，采用免疫组化染色、Westernblotting检测标本MUCl蛋白的表达；术前取患者外周全血400mL，分离外周血单个核细胞（PBMC）并诱导培养树突状细胞（DCs），将DCs分为MUCl组、5μg／mLpolyI：C组、25μg，mLpolyI：C组、50μg／mLpolyI：C组、脂多糖（LPS）组、对照组，分别加入100μL 50μg/mL MUCl、5μg／mL polyI：C、25μg／mLpoly I：C、50μg／mLpolyI：C、50μg／mLLPS和100μL溶剂，培养48h后ELISA法检测上清液中肿瘤坏死因子α（TNF—α）、白介素（IL）-1β、IL-6的浓度；将DCs分为PBS对照组、polyI：C组、MUC1＋polyI：C组，分别加入100μL PBS、50μg，mLpolyI：C、50μG/mLMUCl＋50μg／mL polyI：C，培养48h后流式细胞仪（FCM）检测细胞CD40、CD80、CD83、CD14、组织相容性基因位点抗原DR（HLA—DRl的表达；C57BL／6小鼠24只按随机数字表法分为MUCl组、MUCl＋polyI：C、PBS对照组，每组8只，分别在第1、15天皮下注射1mL50汕卧nLMUCl抗原、50μg／mL CMUCl抗原＋50μg／mLpolyI：C和PBS。注射后7d取各组小鼠脾细胞（CTL）加入到体外培养的小鼠垂体瘤细胞AtT20中，D-乳酸脱氢酶fD．LDH）释放试剂盒检测各组CTL的杀伤活性。结果侵袭性和非侵袭性垂体腺瘤中MUCl表达阳性率分别为90．4％（38／421、20．5％（9／44），MUCl蛋白在侵袭性垂体腺瘤中表达（2．0353±0．0359）高于非侵袭性垂体腺瘤（0．3488±0．0205），差异有统计学意义（B，0．05）；ELISA检测显示MUCl组、5μg／mLpolyI：C组、25μg／mL polyI：C组、50μg／mLpolyI：C组DCs上清液中3种细胞因子的浓度均依次增加，差异有统计学意义�Objective To investigate the MUC 1 protein expression in invasive and non-invasive pituitary adenomas and its induction role in antitumor immune responses. Methods Eighty-six glioma specimens, collected at the resection surgery in our hospitals from March 2009 to December 2014, were used as experimental subjects, including 42 of invasive pituitary adenomas and 44 of non-invasive pituitary adenomas; the MUC1 protein expression was detected by immunohistochemical staining and Western blotting. Before the resection surgery, 400 mL peripheral blood was collected to separate peripheral blood mononuclear cells （PBMCs） and induce dendritic cells （DCs）. （1） The DCs were divided into MUC1 group, 5 μg/mL poly I：C group, 25 μg/mL poly I：C group, 50 μg/mL poly I：C group, lipopolysaccharide （LPS） group and control group, and 100 μL 50 μg/mL MUC1, 5 μg/mL poly I：C, 25 μg/mL poly I：C, 50 μg/mL poly I：C, and 50 μg/mL LPS and 100 μL solvent were given to the above groups; 48 h after the culture, ELISA was employed to detect the concentrations of tumor necrosis factororα（TNFα）, interleukin （IL）-113 and IL-6. （2） The DCs were divided into PBS control group, poly I：C group and MUCl＋poly I：C group, and 100 μL PBS, 50 μg/mL poly I：C and 50 μg/mL MUCI＋50 μg/mL poly I：C were added, respectively; 48 h after the culture, flow cytometry （FCM） was employed to detect the CD40, CD80, CD83, CD14 and histocompatibility genetic locus antigen DR （HLA-DR） expressions. （3） Twenty-four C57BL/6 mice were randomly divided into MUC1 group, MUCl＋poly I：C and PBS control group （n=8）; subcutaneous injection of 1 mL 50 μg/mL MUC1 antigen, 50 μg/mL MUC1 antigen＋50 μg/mL poly I：C and PBS was given to the above three groups on the first and 15th d; 7 d after injection, spleen cytotoxic T lymphocytes （CTLs） from the above three groups were extracted and added into the pituitary adenoma cells AtT20, and D-lactate dehydrogenase （D-LDH） ELISA was used to detect

关 键 词：侵袭性垂体腺瘤 MUC1蛋白 polyI:C 树突状细胞 

分 类 号：R736.4[医药卫生—肿瘤]