Wu Jinding, Wang Delin, Dai Xingliang, Wang Zhongyong, Liu Bing, Lu Zhaohui, Wang Aidong, Dong Jun, Lan Qing, Huang Qiang
Department of Neurosurgery, Second Affiliated Hospital, Soochow University, Suzhou 215004, China.
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Zhonghua Yi Xue Za Zhi. 2014 Sep 23;94(35):2775-80.
Tumor stromal cells have the potential of undergoing malignant transformation induced by glioma stem cells (GSCs) in orthotopic glioma model. The purpose of this study was to explore whether malignant transformation of tumor stromal cells induced by GSCs is dependent on specific local microenvironment.
Human glioma stem/progenitor cell line SU3 transfected with red fluorescent protein (SU3-RFP) gene were implanted into the liver of nude mice with whole-body expressing green fluorescence protein (GFP). Then hepatic tumors were harvested to prepare single cell suspension and analyzed with routine pathological examinations. GFP cells with high proliferative abilities were obtained from the cultivation of single cell suspension. Immortalized glioma stromal cells only expressing GFP and double expressing GFP/RFP were further monocloned with micro-pipetting techniques and under continuous passages. Cell phenotypic analysis and tumorigenicity tests were also performed.
SU3-RFP was transplanted into liver with a tumor formation rate of 83%. Immortalized glioma stromal cells were obtained from re-cultured xenograft tumor tissue. Three monoclonal cell lines B4, B9, B10 were established and proved to be host-derived cells. B4 was found to be a fusion cell co-expressing GFP/RFP and dendritic cell markers CD1 a, CD83 and CD86. Both B9 and B10 were GFP⁺ cells. B9 expressed macrophage markers CD68 and F4/80 while B10 produced fibroblast marker proteins FAP-α, α-SMA and S100. Three cells were all aneuploid with a tumorigenicity rate of 100% in nude mice.
Tumor stromal cells have the potential of malignant transformation in a heterotopic xenograft glioma model. Malignant transformation may also occur outside the central nervous system and contribute to tumor heterogeneity. Further studies are warranted for elucidating the relationship between tumorigenesis, evolution and tumor microenvironment.
在原位胶质瘤模型中，肿瘤基质细胞具有被胶质瘤干细胞（GSCs）诱导发生恶性转化的潜力。本研究旨在探讨GSCs诱导的肿瘤基质细胞恶性转化是否依赖于特定的局部微环境。
将转染了红色荧光蛋白（SU3-RFP）基因的人胶质瘤干/祖细胞系SU3植入全身表达绿色荧光蛋白（GFP）的裸鼠肝脏。然后收获肝肿瘤以制备单细胞悬液，并进行常规病理检查分析。从单细胞悬液培养物中获得具有高增殖能力的GFP细胞。仅表达GFP和同时表达GFP/RFP的永生化胶质瘤基质细胞通过微量移液技术并在连续传代下进一步单克隆化。还进行了细胞表型分析和致瘤性测试。
SU3-RFP移植到肝脏，肿瘤形成率为83%。从重新培养的异种移植肿瘤组织中获得了永生化胶质瘤基质细胞。建立了三个单克隆细胞系B4、B9、B10，并证明是宿主来源的细胞。发现B4是一种共表达GFP/RFP和树突状细胞标志物CD1a、CD83和CD86的融合细胞。B9和B10都是GFP⁺细胞。B9表达巨噬细胞标志物CD68和F4/80，而B10产生成纤维细胞标志物蛋白FAP-α、α-SMA和S100。三种细胞均为非整倍体，在裸鼠中的致瘤率为100%。
在异位异种移植胶质瘤模型中，肿瘤基质细胞具有恶性转化的潜力。恶性转化也可能发生在中枢神经系统之外，并导致肿瘤异质性。有必要进一步研究以阐明肿瘤发生、发展与肿瘤微环境之间的关系。
