Ji X Y, Shi J, Dai X X, Sheng Y J, Xue Y P, Liu J C, Cai H H, Dai X L, Chen Y M, Zhang Y S, Huang Q, Dong J
Department of Neurosurgery, 2nd Affiliated Hospital of Soochow University, Suzhou 215004, China.
Zhonghua Yi Xue Za Zhi. 2018 Nov 6;98(41):3339-3344. doi: 10.3760/cma.j.issn.0376-2491.2018.41.010.
A variety of interstitial cells in tumor microenvironment (TME) based on glioma stem cells(GSC) have the function to promote malignant progression of tumors, but whether these interstitial cells have already undergone malignant transformation and their related molecular characteristics are still poorly understood. Human SU3-RFP glioma stem cells (GSC) stably transfected with red fluorescent protein (RFP) and interstitial cells from enhanced green fluorescent protein (EGFP) transgenic nude mice were co-cultured in vitro. SU3-RFP cells were also inoculated in different tissues of EGFP-Balb/c nude mice. Immortal EGFP(+) cells were monocloned either from co-culture cells in vitro, or from their xenografts in vivo. These immortal EGFP(+) cells were confirmed to bear characteristics of tumor cell via chromosomal analysis and tumorigenicity assay. Related molecular phenotypes of these cells were further detected through RT-PCR, flow cytometry and immunochemistry(IHC) techniques. (1) Two EGFP(+) cell lines were obtained in vitro, and 5 EGFP(+) cell lines were obtained in vivo tumorigenic experiments. Seven EGFP(+) cell lines all have characteristics of self-renewal, heteroploid of chromosomes and 100% tumorigenicity. (2) Cell surface marker analysis showed cell origin of these cell lines were macrophages (tMΦ1 and tMΦ2 ), dendritic cells (tDC1 and tDC2), fibroblasts (tFB), oligodendrocytes (tOG) and BMSC cells (tBMSC), respectively. (3)All of these seven cell lines co-expressed Sca-1 and c-myc, and have Sox-2 or Nanog expression also, which suggest that they may bear molecular characteristics of mesenchymal stem cells or pluripotent stem cells. (1) Tumor stromal cells in TME have undergone malignant transformation, which is related to the tissue remodeling of TME by GSCs, and not limit to the specific type of their parasitic tissues. (2) Tumor cells originated from GSC and tumor interstitial cells, respectively, are two major types of tumor cells with different origins in glioma parenchyma, can not be simply regarded as tumor heterogeneity, transformed interstitial cells of TME may have the potential to serve as new targets for target diagnosis and therapy.
基于胶质瘤干细胞（GSC）的肿瘤微环境（TME）中的多种间质细胞具有促进肿瘤恶性进展的功能，但这些间质细胞是否已经发生恶性转化及其相关分子特征仍知之甚少。将稳定转染红色荧光蛋白（RFP）的人SU3-RFP胶质瘤干细胞（GSC）与增强型绿色荧光蛋白（EGFP）转基因裸鼠的间质细胞进行体外共培养。SU3-RFP细胞也接种到EGFP-Balb/c裸鼠的不同组织中。从体外共培养细胞或其体内异种移植瘤中分离出永生化的EGFP（+）细胞并进行单克隆培养。通过染色体分析和致瘤性试验证实这些永生化的EGFP（+）细胞具有肿瘤细胞的特征。通过RT-PCR、流式细胞术和免疫化学（IHC）技术进一步检测这些细胞的相关分子表型。（1）在体外获得了2株EGFP（+）细胞系，在体内致瘤实验中获得了5株EGFP（+）细胞系。7株EGFP（+）细胞系均具有自我更新、染色体异倍体和100%致瘤性的特征。（2）细胞表面标志物分析表明，这些细胞系的细胞来源分别为巨噬细胞（tMΦ1和tMΦ2）、树突状细胞（tDC1和tDC2）、成纤维细胞（tFB）、少突胶质细胞（tOG）和骨髓间充质干细胞（tBMSC）。（3）这7株细胞系均共表达Sca-1和c-myc，也有Sox-2或Nanog表达，这表明它们可能具有间充质干细胞或多能干细胞的分子特征。（1）TME中的肿瘤基质细胞发生了恶性转化，这与GSCs对TME的组织重塑有关，且不限于其寄生组织的特定类型。（2）分别起源于GSC和肿瘤间质细胞的肿瘤细胞是胶质瘤实质中两种不同起源的主要肿瘤细胞类型，不能简单地视为肿瘤异质性，TME中转化的间质细胞可能有潜力成为靶向诊断和治疗的新靶点。
