[一种新的人胶质瘤细胞系的建立及其生物学特性分析]

[Establishment of a new human glioma cell line and analysis of its biological characteristics].

作者信息

Chen Guilin, Li Yanyan, Xie Xueshun, Chen Jinming, Wu Tingfeng, Li Xuetao, Wang Hangzhou, Zhou Youxin, Du Ziwei

机构信息

Neurosurgery & Brain and Nerve Research Laboratory, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.

Email:

出版信息

Zhonghua Zhong Liu Za Zhi. 2015 Feb;37(2):84-90.

PMID:25975682

Abstract

OBJECTIVE

To establish a new glioma cell line and analyze its biological characteristics, and to provide a useful cellular tool with new features for cancer research.

METHODS

Glioma tissue was taken from surgical specimen clinical of a clinical patient. Primary culture was carried out, and a cell line (SHG139) was established after 10 passages. Immunofluorescence staining was performed to detect the expression of proteins, and cell proliferation and cycle were detected by flow cytometry method (FCM). The biological characteristics of SHG139 cells were detected by chromosome karyotype analysis. SHG139s glioma cells derived from SHG139 glioma cell line were cultured with neural stem cell medium. Then stem cell markers were determined. SHG139s cells were induced with serum-containing medium, and their expression of A2B5, GFAP, β-III tubulin, and GalC was detected. Intracranial xenograft tumor of both SHG139 glioma cells and SHG139s glioma stem cell spheres was generated in rats.

RESULTS

The expressions of A2B5, GalC, GFAP, S-100, and vimentin in the 20 and 60 passages of SHG139 cells were positive, consistent with the immunohistochemical results and pathological features. SHG139 cells proliferated significantly within 24 h after subculture, and their total number of chromosomes was 68 and mostly multiploid. They were positive for A2B5 (84.12±9.96)%, nestin (73.86±5.01)%, and NG2 (73.37±2.09)%. SHG139s cells were induced, and the ratio of positive cells of GFAP, β-III tubulin and GalC was (92.89±2.24)%, (64.85±4.09)% and (33.57±4.14)%, respectively.

CONCLUSIONS

SHG139 is an astroglioma cell line, from which SHG139s cells can be successfully obtained by culture with NSCM. SHG139s cells are of A2B5(+)/CD133(-) GSCs subgroup cells, with potentials of self-renewal and multi-directional differentiation. Compared with the intracranial SHG139 xenograft tumor, the intracranial SHG139s xenograft tumor is more malignant and aggressive.

摘要

目的

建立一种新的胶质瘤细胞系并分析其生物学特性，为癌症研究提供具有新特性的有用细胞工具。

方法

从一名临床患者的手术标本中获取胶质瘤组织，进行原代培养，传代10次后建立细胞系（SHG139）。采用免疫荧光染色检测蛋白表达，通过流式细胞术（FCM）检测细胞增殖和周期。通过染色体核型分析检测SHG139细胞的生物学特性。将源自SHG139胶质瘤细胞系的SHG139s胶质瘤细胞用神经干细胞培养基培养，然后测定干细胞标志物。用含血清培养基诱导SHG139s细胞，检测其A2B5、GFAP、β-III微管蛋白和半乳糖脑苷脂（GalC）的表达。在大鼠体内生成SHG139胶质瘤细胞和SHG139s胶质瘤干细胞球的颅内移植瘤。

结果

SHG139细胞传代20代和60代时，A2B5、GalC、GFAP、S-100和波形蛋白的表达均为阳性，与免疫组化结果及病理特征相符。SHG139细胞传代培养后24小时内增殖明显，其染色体总数为68条，大多为多倍体。它们的A2B5阳性率为（84.12±9.96）%，巢蛋白阳性率为（73.86±5.01）%，NG2阳性率为（73.37±2.09）%。诱导SHG139s细胞后，GFAP、β-III微管蛋白和GalC的阳性细胞比例分别为（92.89±2.24）%、（64.85±4.09）%和（33.57±4.14）%。