人脑胶质瘤干细胞-裸小鼠连续皮下移植变成纤维肉瘤的报告  

Series subcutaneous transplantation of human glioma cells led to transformation of fibrosarcoma in green fluorescence protein nude mice

作　　者：戴纯刚[1] 刘兵[1] 马加威 代兴亮[2] 芮琴[2] 贾敬云 谢涛[1] 马笑宇[1] 陆朝晖[3] 董军[3] 兰青[3] 黄强[3] 

机构地区：[1]苏州大学附属第二医院高新区医院神经外科,苏州215151 [2]苏州大学附属第二医院实验中心,苏州215004 [3]苏州大学附属第二医院神经外科,苏州215004

出　　处：《中华神经医学杂志》2016年第3期231-236,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81172400、81272799、81472739）;苏州科技计划项目（sYS201036）

摘　　要：目的探讨源于人脑胶质瘤干细胞的绿色荧光裸小鼠皮下移植瘤组织，经长期鼠间传代后演变成纤维肉瘤的可能性。方法取浓度为1×10^6个/0.5 mL PBS的SU3胶质瘤干细胞，接种于纯合子NC-C57BL6J-增强型绿色荧光蛋白（EGFP）＋小鼠皮下，致瘤后将1 mm3大小的移植瘤组织经套管针行同品系鼠间传代移植，观察移植瘤的增殖特征。取初代移植瘤和各鼠间传代的肿瘤组织进行冰冻切片，荧光显微镜下观察肿瘤组织中发绿色荧光的细胞；HE染色肿瘤组织；免疫组织化学染色检测肿瘤组织乏氧诱导因子α（HIF-α）、肿瘤坏死因子α（TNF-α）、胶质纤维酸性蛋白（GFAP）和成纤维细胞活化蛋白（FAP）的表达。结果前3代移植致瘤率是100%，肿瘤大小的个体差异较大，4-8代以后差距逐渐缩小，且增殖规律符合人脑胶质瘤裸小鼠移植模型特征。随着移植瘤传代次数增加，EGFP＋细胞在肿瘤组织中所占比例逐渐增加。HE染色显示肿瘤组织坏死、出血多见，血管丰富，核大，深染、异型明显，向肌肉间隙呈广泛侵袭性生长。免疫组织化学染色显示TNF-α＋细胞、HIF-α＋细胞广泛分布于各代移植瘤组织中，无GFAP＋细胞，90%的细胞FAP呈阳性表达，诊断为纤维肉瘤。结论源于人脑胶质瘤干细胞的裸小鼠皮下移植瘤组织经长期鼠间传代后会变成鼠源性纤维肉瘤，说明肿瘤细胞在寄生的宿主组织重构过程中能诱导成纤维间质细胞的恶性转化。Objective To investigate the possibilities of fibrosarcoma transformation in long term serial transplantation of human glioma cells subcutaneously in nude mice with whole body green fluorescence expression. Methods Firstly, 1× 10^6 human brain glioma stem cells SU3 were seeded in NC-C57BL6J-enhanced green fluorescent protein nude mice subcutaneously, and tumors were harvested and cut into pieces （1 mm^3/piece） and passaged in vivo continuously; after formation of transplanted tumors, tumor proliferation was observed; histological changes （cells with green fluorescence） were observed under fluorescence microscope; HE staining was performed on the rumor tissues; cal staining was used to detect the expressions of hypoxia-inducible factorα （HIF-α）, tumor necrosis factor α （TNF-α） and fibroblast activated protein （FAP）. Results During the first three generations of transplanted tumors, tumorigenic rate was 100%, but the individual difference of tumor size was obvious; however, tumor size differences decreased when being passaged in vivo after four to eight generations. Tumorigenicity rate was 100% in EGFP nude mice. EGFP positive cells of original host stromal cell ratio during continuous passages in vivo increased gradually. HE staining indicated tumor tissue necrosis and bleeding, abundant blood vessel, large nucleus, deep dyed and abnormal shaped cells, and invasive growth to the muscle gap; immunohistochemical staining showed that the TNF＋ and HIF-α＋ cells widely distributed at the transplanted tumors of different generations, and no GFAP＋ cells were noted; 90% cells had FAP positive expression; fibrosarcomas could be verified. Conclusion Nude mice transplanted tumors originated from human brain glioma stem cells could turn into fibrosarcoma after long term inter-mice passage, which suggests that tumor cells in the host tissue remodeling via parasitic processes can induce malignant transformation of fibrous stroma cells.

关 键 词：神经胶质瘤 荧光裸小鼠 肿瘤移植 间质细胞恶性转化 

分 类 号：R739.41[医药卫生—肿瘤]