Chang F, Li R, Noon K, Gage D, Ladisch S
Glycobiology Program, Children's Research Institute, Washington, DC, USA.
Glycobiology. 1997 Jun;7(4):523-30. doi: 10.1093/glycob/7.4.523.
To establish a model system for the study of ganglioside metabolism of the human brain tumor, medulloblastoma, we have chemically characterized the gangliosides of the Daoy cell line. These cells contain a high concentration of gangliosides (143 +/- 13 nmol LBSA/10(8) cells). The major species have been structurally confirmed to be GM2 (65.9%), GM3 (13.0%), and GD1a (10.3%). Isolation of individual gangliosides homogeneous in both carbohydrate and ceramide moieties by reversed-phase HPLC and analysis by negative-ion fast atom bombardment collisionally activated dissociation tandem mass spectrometry have allowed us to unequivocally characterize ceramide structures. In the case of GM2, 10 major ceramide subspecies were identified: d18:1-hC16:0, d18:1-C16:0, d18:0-C16:0, d18:1-C18:0, d18:1-C20:0, d18:1-C22:0, d18:2-C24:1, d18:1-C23:1, d18:1-C24:1, and d18:1-C24:0. Taken together with previous studies, these findings in buman medulloblastoma cells support the view that high expression and marked heterogeneity of ceramide structure are general characteristics of tumor gangliosides, molecules which are shed by the tumor cells and which are biologically active in vivo.
为建立用于研究人脑肿瘤髓母细胞瘤神经节苷脂代谢的模型系统,我们已对道氏(Daoy)细胞系的神经节苷脂进行了化学表征。这些细胞含有高浓度的神经节苷脂(143±13 nmol LBSA/10⁸ 个细胞)。主要种类经结构确认为GM2(65.9%)、GM3(13.0%)和GD1a(10.3%)。通过反相高效液相色谱法分离碳水化合物和神经酰胺部分均同质的单个神经节苷脂,并通过负离子快原子轰击碰撞激活解离串联质谱法进行分析,这使我们能够明确表征神经酰胺结构。就GM2而言,鉴定出10种主要的神经酰胺亚类:d18:1 - hC16:0、d18:1 - C16:0、d18:0 - C16:0、d18:1 - C18:0、d18:1 - C20:0、d18:1 - C22:0、d18:2 - C24:1、d18:1 - C23:1、d18:1 - C24:1和d18:1 - C24:0。结合先前的研究,这些在人髓母细胞瘤细胞中的发现支持以下观点:神经酰胺结构的高表达和显著异质性是肿瘤神经节苷脂的普遍特征,肿瘤细胞释放的这些分子在体内具有生物活性。