Zhang Lei, Yamane Tetsu, Satoh Eiji, Amagasaki Kenichi, Kawataki Tomoyuki, Asahara Takayuki, Furuya Koro, Nukui Hideaki, Naganuma Hirofumi
Department of Neurosurgery, University of Yamanashi Faculty of Medicine, Yamanashi, Japan.
Neuropathology. 2005 Jun;25(2):136-43. doi: 10.1111/j.1440-1789.2005.00601.x.
Five malignant glioma cell lines (YMG1, 2, 3, 4, and 5) were established from surgical specimens obtained from patients with glioblastoma or anaplastic astrocytoma, and these lines were partially characterized. Three glioma cell lines (YMG1, 3, and 5) were weakly positive for GFAP by Western blot analysis and two cell lines were negative. S-100 protein was positive in all glioma cell lines. The expression of p53, p16, p15, cyclin-dependent kinase 4 (CDK4), and EGF receptor (EGFR) proteins was examined by Western blotting. YMG1 and 2 cell lines showed accumulation of p53 protein and loss of p16 and p15 expression. YMG3 and 4 showed accumulation of p53 protein and expression of p16 and p15 proteins. YMG5 revealed weak expression of p53 protein, suggesting wild-type p53, and loss of p16 and p15 expression. All cell lines expressed various levels of CDK4 protein. YMG1, 2, and 3 showed higher EGFR protein expression and YMG4 and 5 showed lower EGFR expression compared to U251 glioblastoma cells, which express high levels of EGFR. Fluorescence in situ hybridization analysis for EGFR gene expression did not show any amplification in the glioma cell lines. Immunohistochemical studies revealed that the patterns of p53 and EGFR expressions in the original tumor tissues were mostly correlated with those in the malignant glioma cell lines. These results suggest that the characteristics of p53 and EGFR expression in the malignant glioma cell lines were passed over from the original tumor tissues. These newly established malignant glioma cell lines can be used for further analysis of the mechanisms of tumor growth and progression.
从胶质母细胞瘤或间变性星形细胞瘤患者的手术标本中建立了5种恶性胶质瘤细胞系（YMG1、2、3、4和5），并对这些细胞系进行了部分特征分析。通过蛋白质免疫印迹分析，3种胶质瘤细胞系（YMG1、3和5）的胶质纤维酸性蛋白（GFAP）呈弱阳性，2种细胞系为阴性。S-100蛋白在所有胶质瘤细胞系中均呈阳性。通过蛋白质免疫印迹法检测了p53、p16、p15、细胞周期蛋白依赖性激酶4（CDK4）和表皮生长因子受体（EGFR）蛋白的表达。YMG1和2细胞系显示p53蛋白积累，p16和p15表达缺失。YMG3和4显示p53蛋白积累以及p16和p15蛋白表达。YMG5显示p53蛋白弱表达，提示为野生型p53，同时p16和p15表达缺失。所有细胞系均表达不同水平的CDK4蛋白。与高表达EGFR的U251胶质母细胞瘤细胞相比，YMG1、2和3显示出更高的EGFR蛋白表达，而YMG4和5显示出较低的EGFR表达。对EGFR基因表达进行的荧光原位杂交分析未显示胶质瘤细胞系中有任何扩增。免疫组织化学研究表明，原发肿瘤组织中p53和EGFR的表达模式大多与恶性胶质瘤细胞系中的表达模式相关。这些结果表明，恶性胶质瘤细胞系中p53和EGFR的表达特征是从原发肿瘤组织传递而来的。这些新建立的恶性胶质瘤细胞系可用于进一步分析肿瘤生长和进展的机制。
