丝裂霉素诱导的肝癌细胞凋亡致敏树突状细胞(英文)  

Sensitized dendritic cells with apoptotic hepatoma cells induced by mitomycin

作　　者：吴刚[1] 韩本立[2] 裴雪涛[3] 

机构地区：[1]解放军第三军医大学大坪医院肝胆外科,重庆市400042 [2]解放军第三军医大学西南医院肝胆外科,重庆市400038 [3]解放军军事医学科学院放射医学研究所实验血液研究室,北京市100850

出　　处：《中国临床康复》2006年第32期190-192,i0001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家杰出青年科学基金资助项目(39825111)~~

摘　　要：目的:建立丝裂霉素诱导癌细胞凋亡致敏从人外周血诱导扩增的树突状细胞的方法。设计:以癌细胞凋亡致敏树突状细胞为观察对象的随机对照实验。单位:解放军第三军医大学大坪医院野战外科研究所,解放军军事医学科学院放射医学研究所。材料:实验于1998-04/1999-05在解放军军事医学科学院放射医学研究所完成。细胞株为QBC939胆管癌细胞株,药物为抗肿瘤药物丝裂霉素。方法:从正常人外周血分离获得单核细胞,加入50μg/L重组人粒细胞集落刺激因子,1000U/mL白细胞介素4,隔天一次,共4次,培养第3天,加入丝裂霉素诱导凋亡的胆管癌细胞,再继续体外培养4d后,用树突细胞富集柱收集树突状细胞。主要观察指标:①培养的树突状细胞的鉴定。②树突状细胞和坏死胆管癌细胞以及正常培养的胆管癌细胞共培养,观察其吞噬凋亡小体负载抗原。③检测不同密度的树突状细胞(1×103/孔,5×103/孔,1×104/孔)的免疫刺激活性及负载抗原后的免疫刺激活性,以单核细胞作对照组。结果:①培养、扩增得到的树突状细胞高表达共刺激分子B7和CDla,表面具有典型不规则突起。②丝裂霉素诱导癌细胞凋亡形成的凋亡小体被树突状细胞捕捉、吞噬。③负载抗原的树突状细胞其激发同种异体T淋巴细胞增殖的能力进一步增强。结论:丝裂霉素诱导癌细胞凋亡可以致敏重组人粒细胞集落刺激因子加重组人白细胞介素4从人外周血单核细胞诱导、扩增出的树突状细胞,树突状细胞可以有效提呈凋亡胆管癌细胞的抗原,可望成为有效的肿瘤抗原致敏树突状细胞的新途径。BACKGROUND： Dendritic cells play an important role in antigen present in vivo, and the mechanism of tumor cells in escaping the antigen presentation of dendritic cells existed in the patients with tumor. OBJECTIVE： To sensitize dendritic cells from human peripheral blood with apoptotic hepatoma cells induced by mitomycin. DESIGN： A randomized control trial by taking apoptotic hepatoma cell sensitized dendritic cells as the observed subjects. SETTINGS： Institute of Field Surgery, Daping Hospital, the Third Military Medical University of Chinese PLA; Institute of Radiation Medicine, Chinese PLA Academy of Military Medical Sciences. MATERIALS： The experiments were carried out in the Institute of Radiation Medicine, Chinese PLA Academy of Military Medical Sciences from April 1998 to May 1999. Tile cell strain was the QBC939 bile duct cancer cell strain, and mitomycin was used as the antitumor drug. METHODS： Mononuclear cells were isolated from the peripheral blood of normal people, 50μg/L granulocyte：macrophage colony stimulating factor （GM-CSF） and 1 000 U/mL interleukin-4 （IL-4） were added, once every other day for 4 times. On the 3^th day of culture, the apoptotic bile duct cancer cells induced by mitomycin was added, and then cultured in vitro for 4 days, finally the dendritic cells were collected. MAIN OUTCOME MEASURES： ① The identification of the cultured dendritic cells was observed; ② The dendritic cells were co-cultured with necrotic and normally cultured bile duct cancer cells respectively, and the phagocytized apoptotic body loaded antigens were observed; ③ The immunostimulatory activity of dendritic cells （1 ×10^3, 5×10^3 and 1×10^4/well） and that after loaded by antigen were detected, and the mononuclear cells were taken as controls. RESULTS： ② The cultured and amplified dendritic cells expressed high levels of costimulatory molecules of CDla and B7, and there were typical irregular processes on the surface. ② The tumor cells formed apoptotic bodies wh

关 键 词：丝裂霉素 树突细胞 单核细胞 粒细胞巨噬细胞集落刺激因子 白细胞介素4 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]