Ariga T, Blaine G M, Yoshino H, Dawson G, Kanda T, Zeng G C, Kasama T, Kushi Y, Yu R K
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298, USA.
Biochemistry. 1995 Sep 12;34(36):11500-7. doi: 10.1021/bi00036a024.
We have studied the glycosphingolipid composition in an F-11 neuroblastoma cell line originated from hybridization of a mouse neuroblastoma cell line (N18TG-2) with rat dorsal root ganglion cells. The total lipid-bound glucose of F-11 cells was estimated to be 0.28 micrograms/mg of protein and the total lipid-bound sialic acid was 0.82 micrograms/mg of protein. The major neutral glycosphingolipids were Gb4 (37% of the total neutral glycosphingolipids), Gb3 (15%), LacCer (21%), and GlcCer (15%). The major gangliosides were found to be GM3 (37% of the total gangliosides), GD3 (27%), O-acetylated GD3 (18%), and GD1a (4%), with trace amounts of GD2. The unusually high concentration of O-acetylated GD3 is consistent with its putative role as a tumor marker. Immunocytochemical localization studies of GD3 and O-acetylated GD3, examined by mouse monoclonal antibodies R24 and D1.1, respectively, revealed that the cell bodies and processes were all positively stained. To elucidate the role of O-acetylated GD3 in tumorigenesis, we transfected F-11 cells with the O-acetylesterase gene from influenza C virus. Compared with the original cell line, the transfected cells showed a dramatic increase in the level of GD3 (150% of that in the control cells) and a significant decrease of the concentration of O-acetylated GD3 (27% of control cells). In addition, the transfected F-11 cells exhibited a morphology different from the parental cells with enlarged cell bodies and elongated neurites. We conclude that alteration of ganglioside composition, particularly the expression of GD3 and O-acetylated GD3, may be associated with the morphological changes observed in this cell line.(ABSTRACT TRUNCATED AT 250 WORDS)
我们研究了一种F-11神经母细胞瘤细胞系中的糖鞘脂成分，该细胞系源自小鼠神经母细胞瘤细胞系（N18TG-2）与大鼠背根神经节细胞的杂交。F-11细胞中与脂质结合的葡萄糖总量估计为0.28微克/毫克蛋白质，与脂质结合的唾液酸总量为0.82微克/毫克蛋白质。主要的中性糖鞘脂是Gb4（占总中性糖鞘脂的37%）、Gb3（15%）、乳糖神经酰胺（21%）和葡萄糖神经酰胺（15%）。主要的神经节苷脂被发现是GM3（占总神经节苷脂的37%）、GD3（27%）、O-乙酰化GD3（18%）和GD1a（4%），还有微量的GD2。O-乙酰化GD3异常高的浓度与其作为肿瘤标志物的假定作用一致。分别用小鼠单克隆抗体R24和D1.1检测GD3和O-乙酰化GD3的免疫细胞化学定位研究表明，细胞体和突起均呈阳性染色。为了阐明O-乙酰化GD3在肿瘤发生中的作用，我们用丙型流感病毒的O-乙酰酯酶基因转染了F-11细胞。与原始细胞系相比，转染后的细胞GD3水平显著增加（为对照细胞的150%），O-乙酰化GD3浓度显著降低（为对照细胞的27%）。此外，转染后的F-11细胞呈现出与亲代细胞不同的形态，细胞体增大，神经突伸长。我们得出结论，神经节苷脂组成的改变，特别是GD3和O-乙酰化GD3的表达，可能与该细胞系中观察到的形态变化有关。（摘要截短于250字）
