一种新的转化为A2B5(+)干细胞的人胶质瘤细胞系的生物学特性
Biological characteristics of a new human glioma cell line transformed into A2B5(+) stem cells.
作者信息
Li Yanyan, Wang Hangzhou, Sun Ting, Chen Jinming, Guo Lingchuan, Shen Haitao, Du Ziwei, Zhou Youxin
机构信息
Department of Neurosurgery and Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, Jiangsu, China.
Department of Pathology, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, Jiangsu, China.
出版信息
Mol Cancer. 2015 Apr 2;14:75. doi: 10.1186/s12943-015-0343-z.
OBJECTIVE
The new glioma cell line SHG-139 was established and its phenotype, tumorigenicity, pathological characteristics, derived stem cells SHG139S were studied.
METHODS
Immunohistochemistry was used to assess expressions in the patient and mouse tumor tissues, SHG-139 and SHG-139S. Primary SHG-139 culture was performed, cell proliferation, cell cycle and genetic characteristics were assessed. MiRNA (Micro RNA) and LncRNA (Long non-coding RNA) microarray was performed.
RESULTS
We found that the glioma tissue was positive for A2B5 (Glial precursors ganglioside), GFAP (Glial fibrillary acidic protein), S-100 (Acid calcium bingding protein), VEGF (Vascular endothelial growth factor), VEGFR (Vascular endothelial growth factor receptor) and negative for Ki-67 (Nuclcar- associated antigen). SHG-139 proliferated significantly within 24h; its total number of chromosomes was 68; ratios of SHG-139 and SHG-139S cells in G1 phase were highest. SHG-139 cells were positive for A2B5, GalC (Galactocerebrosides), GFAP, S-100 and Vimentin, while SHG-139S cells were positive for A2B5, Nestin, and NG2 (Neuron-glia antigen2), and negative for Vimentin and IDHR132H (Isocitrate dehydrogenase); cells rarely stained for CD133 (Cluster of differentiation133). SHG-139 intracranial xenografts expressed GFAP, but no overt oligodendroglioma was observed. In SHG-139S xenografts, GFAP and S-100 were expressed, while CD133 was not detected; a few A2B5(+) cells were found at tumor edges, and typical oligodendroglioma were obtained. In addition, SHG-139S xenograft tumors were more aggressive than those of SHG-139. Anti-mouse CD31 (Cluster of differentiation31) staining revealed murine vessels at the border between xenograft tumor and normal brain tissue; Anti-human CD34 (Cluster of differentiation34) staining was negative. Biochip technology of SHG139S showed several miRNA and lncRNA were differently expressed in SHG139 and SHG139S.
CONCLUSIONS
SHG-139 was an astroglioma cell line which yielded stem cells SHG-139S. SHG-139S cells constituted an A2B5(+)/CD133(-) GSC subgroup.
目的
建立新的胶质瘤细胞系SHG - 139，并研究其表型、致瘤性、病理特征以及其衍生的干细胞SHG139S。
方法
采用免疫组织化学法评估患者及小鼠肿瘤组织、SHG - 139和SHG - 139S中的表达情况。进行SHG - 139原代培养，评估细胞增殖、细胞周期及遗传特征。进行miRNA（微小RNA）和LncRNA（长链非编码RNA）芯片检测。
结果
我们发现胶质瘤组织中A2B5（神经胶质前体细胞神经节苷脂）、GFAP（胶质纤维酸性蛋白）、S - 100（酸性钙结合蛋白）、VEGF（血管内皮生长因子）、VEGFR（血管内皮生长因子受体）呈阳性，而Ki - 67（核相关抗原）呈阴性。SHG - 139在24小时内显著增殖；其染色体总数为68条；SHG - 139和SHG - 139S细胞在G1期的比例最高。SHG - 139细胞中A2B5、GalC（半乳糖脑苷脂）、GFAP、S - 100和波形蛋白呈阳性，而SHG - 139S细胞中A2B5、巢蛋白和NG2（神经胶质抗原2）呈阳性，波形蛋白和IDHR132H（异柠檬酸脱氢酶）呈阴性；细胞很少表达CD133（分化簇133）。SHG - 139颅内异种移植瘤表达GFAP，但未观察到明显的少突胶质细胞瘤。在SHG - 139S异种移植瘤中，GFAP和S - 100表达，而未检测到CD133；在肿瘤边缘发现少数A2B5(+)细胞，并获得典型的少突胶质细胞瘤。此外，SHG - 139S异种移植瘤比SHG - 139的更具侵袭性。抗小鼠CD31（分化簇31）染色显示异种移植瘤与正常脑组织边界处有小鼠血管；抗人CD34（分化簇34）染色为阴性。SHG139S的生物芯片技术显示几种miRNA和lncRNA在SHG139和SHG139S中表达不同。
结论
SHG - 139是一种星形胶质瘤细胞系，可产生干细胞SHG - 139S。SHG - 139S细胞构成一个A2B5(+)/CD133(-)的胶质瘤干细胞亚群。
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