[SHG-44胶质瘤干细胞球的表型及其异种移植瘤的病理特征]

[Phenotype of SHG-44 glioma stem cell spheres and pathological characteristics of their xenograft tumors].

作者信息

Wu Ting-feng, Chen Jin-ming, Chen San-song, Chen Gui-lin, Wei Yong-xin, Xie Xue-shun, Du Zi-wei, Zhou You-xin

机构信息

Department of Neurosurgery, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.

Department of Neurosurgery, the First Affiliated Hospital of Soochow University, Suzhou 215006, China. Email:

出版信息

Zhonghua Zhong Liu Za Zhi. 2013 Oct;35(10):726-31.

PMID:24378091

Abstract

OBJECTIVE

To study the phenotype and tumorigenicity of SHG-44 glioma stem cell spheres and the pathological characteristics of their xenograft tumors.

METHODS

SHG-44 glioma cells were cultured under neural stem cell medium and glioma stem cell spheres were collected. Immunocytochemistry was used to dectet the expression of CD133, nestin, A2B5, vimentin, VEGFR-2 and IDH R132H. Cell spheres were induced using serum-containing medium, and the expression of CD133, nestin, vimentin, GFAP, β-III tubulin and GalC in the cell spheres were detected. The expression of CD133, nestin, VEGFR-2, GFAP, S-100 and CD34 in the intracranial xenograft tumor tissues was detected using immunohistochemistry. The pathological characteristics of orthotopic xenograft tumors generated from the SHG-44 glioma cells and SHG-44 glioma stem cell spheres were compared.

RESULTS

SHG-44 glioma stem cell spheres were collected successfully after cultured under neural stem cell medium. The ratio of CD133(+) cells in the passage 10 SHG-44 glioma stem cell spheres was (71.63 ± 5.92)%, significantly higher than that in the SHG-44 glioma cells [(1.95 ± 1.45)%]. Immunocytochemistry showed that in the SHG-44 glioma cell spheres, the ratio of nestin(+) cells was (84.06 ± 7.58)%, vimentin(+) cells (29.11 ± 3.44)%, VEGFR 2(+) cells (64.44 ± 3.69)%, and A2B5(+) cells (14.08 ± 2.19)%. A subpopulation of cells with mutation of IDH R132H was detected harboring in the SHG-44 glioma cell spheres. After induction of differentiation with serum-containing medium, the ratio of CD133(+) cells was (1.89 ± 1.27)%, nestin(+) cells (6.67 ± 2.75)%, vimentin(+) cells (93.75 ± 2.95)%, GFAP (+) cells (91.33 ± 4.75)%, β-III tubulin(+) cells (82.36 ± 4.02)%, and GalC(+) cells (8.92 ± 3.19)%. Immunohistochemistry showed positive expression of GFAP, S-100, VEGFR-2, and negative of CD133 and nestin in the orthotopic xenograft tumors. A very small amount of human-specific CD34 cells formed a tubular structure. Compared with the SHG-44 glioma cell-formed xenograft tumor, the SHG-44 glioma stem cell-formed xenograft tumor exhibited a higher local invasiveness.

CONCLUSIONS

SHG-44 glioma cell spheres are successfully collected after cultured under neural stem cell medium. They belong to the CD133(+)A2B5(-) GSC subpopulation, highly expressing VEGFR-2, possess the ability of both self-renewal and multi-directional differentiation, and may participate in the formation of vasculogenic mimicry.

摘要

目的

研究SHG-44胶质瘤干细胞球的表型、致瘤性及其异种移植瘤的病理特征。

方法

将SHG-44胶质瘤细胞在神经干细胞培养基中培养，收集胶质瘤干细胞球。采用免疫细胞化学法检测CD133、巢蛋白、A2B5、波形蛋白、血管内皮生长因子受体2（VEGFR-2）和异柠檬酸脱氢酶（IDH）R132H的表达。用含血清培养基诱导细胞球分化，检测细胞球中CD133、巢蛋白、波形蛋白、胶质纤维酸性蛋白（GFAP）、β-Ⅲ微管蛋白和半乳糖脑苷脂（GalC）的表达。采用免疫组织化学法检测颅内异种移植瘤组织中CD133、巢蛋白、VEGFR-2、GFAP、S-100和CD34的表达。比较SHG-44胶质瘤细胞和SHG-44胶质瘤干细胞球原位异种移植瘤的病理特征。

结果

在神经干细胞培养基中培养后成功收集到SHG-44胶质瘤干细胞球。第10代SHG-44胶质瘤干细胞球中CD133(+)细胞比例为(71.63±5.92)%，显著高于SHG-44胶质瘤细胞[(1.95±1.45)%]。免疫细胞化学显示，SHG-44胶质瘤细胞球中巢蛋白(+)细胞比例为(84.06±7.58)%，波形蛋白(+)细胞(29.11±3.44)%，VEGFR 2(+)细胞(64.44±3.69)%，A2B5(+)细胞(14.08±2.19)%。在SHG-44胶质瘤细胞球中检测到一小部分IDH R132H突变细胞亚群。用含血清培养基诱导分化后，CD133(+)细胞比例为(1.89±1.27)%，巢蛋白(+)细胞(6.67±2.75)%，波形蛋白(+)细胞(93.75±2.95)%，GFAP(+)细胞(91.33±4.75)%，β-Ⅲ微管蛋白(+)细胞(82.36±4.02)%，GalC(+)细胞(8.92±3.19)%。免疫组织化学显示，原位异种移植瘤中GFAP、S-100、VEGFR-2呈阳性表达，CD133和巢蛋白呈阴性表达。极少量人特异性CD34细胞形成管状结构。与SHG-44胶质瘤细胞形成的异种移植瘤相比，SHG-44胶质瘤干细胞形成的异种移植瘤具有更高的局部侵袭性。