在SU3-RFP／GFP模型中观察胶质瘤间质细胞恶性转化  

Observation of malignant transformation of the stroma cells in SU3-RFP/GFP models

作　　者：代兴亮[1] 戴纯刚[2] 贾敬云 谢涛[2] 刘兵[2] 马笑宇[2] 陆朝晖[1,2] 王爱东[1] 董军[1] 兰青[1] 黄强[1] 

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

出　　处：《中华神经医学杂志》2014年第8期757-762,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81071766、81172400、81272799）;江苏省自然科学基金（BK2010227）;苏州市应用基础研究计划（SYS201036）;苏州市科技兴卫基金（SYS201269）

摘　　要：目的 通过建立小鼠移植瘤模型及荧光蛋白示踪技术,探讨肿瘤间质细胞恶性转化的相关特性. 方法 将转染红色荧光蛋白（RFP）的人脑胶质瘤干/祖细胞SU3接种于表达绿色荧光蛋白（GFP）的裸小鼠皮下,致瘤后取移植瘤组织再培养,克隆永生化的间质细胞,分析这些细胞体外生长特性、DNA合成能力、细胞染色体特征以及在BALB/c无胸腺裸小鼠中的致瘤情况. 结果 SU3-RFP干/祖细胞接种GFP裸小鼠的致瘤率为100％（12/12）.移植瘤组织再培养的细胞群中,克隆到了ihSTC-H8、ihSTC-F8和ihSTC-S2（简称H8、F8和S2）3株具有永生化特征且表达GFP和成纤维细胞标志蛋白——成纤维细胞活化蛋白（FAP）、S100钙结合蛋白A4（S100A4）的细胞,并且具备异常高的DNA合成能力和超5倍体的染色体核型（H8、F8和S2染色体条数分别为110.58±6.51、106.17±6.36和102.58±6.94）,在BALB/c无胸腺裸小鼠中达到100％（9/9）的致瘤率. 结论 异位生长在宿主皮下的SU3-RFP胶质瘤干/祖细胞具有诱导表达S100A4和FAP的间质成纤维细胞恶性转化的潜能.Objective To investigate the malignant transformation of the stroma cells via transplanting the glioma stem/progenitor cells labeled by red fluorescence protein gene （RFP） into the subcutaneous tissues of the transgenic nude mice expressed green fluorescence protein （GFP）.Methods Previously established human glioma stem/progenitor cells SU3 were transfected with red fluorescence protein gene （SU3-RFP）,and transplanted subcutaneously to the transgenic GFP athymic nude mice.Then,the xenograft tumor was re-cultured,and the immortalized stromal cells expressing GFP were sorted and cloned.Further analyses of in vitro growth characteristics,DNA content,chromosome banding and tumorigenesis abilities of the immortalized stromal cells were performed.Results Tumorigenic rate of the SU3-RFP stem cells transplanted to the subcutaneous of the GFP nude mice was 100% （12/12）.Three immortalized stromal cell lines （named H8,F8 and S2） expressing GFP,enjoyed fibroblast activation protein （FAP） and S100 calcium-binding protein A4 （S100A4） positive expressions,were cloned,and unusually high DNA content and telocentric （murine） chromosome with ultra-pentaploid （H8,F8 and S2 were 110.58±6.51,106.17±6.36 and 102.58±6.94） were observed; the tumorigenesis rate of the immortalized stromal cells in BALB/c athymic nude mice was 100% （9/9）.Conclusion SU3-RFP cells transplanted to the host＇ s subcutaneous tissues have the potential to induce malignant transformation of the stroma cells,which can become S100A4 and FAP positive fibroblasts.

关 键 词：荧光蛋白示踪 肿瘤干细胞 肿瘤移植 间质细胞 转化 

分 类 号：R739.41[医药卫生—肿瘤]