Regulation of cell cycle progression by ganglioside-a novel strategy against malignant glioma

为了研究神经节苷脂是否调节神经胶质瘤细胞的生长，将最简单的神经节苷脂GM3通过枕大池注入神经胶质瘤细胞培养和荷瘤大鼠体内。(1)用GM3培养4株原代胶质瘤细胞和4株胶质瘤细胞。四甲基偶氮唑盐比色法显示，浓度为20~200微摩尔的GM3对胶质瘤细胞的增殖有抑制作用(P&lt；0.05)。抑制作用依赖于GM3的浓度。(2)将C6胶质瘤细胞接种于大鼠脑膜腔内，建立可用于体内分析的大鼠胶质瘤模型。GM3治疗组(n=20)在胶质瘤细胞接种后10d存活22.0天，对照组(n=15)存活19.5天(P&lt；0.05)。GM3给药大鼠未检测到急性毒性。(3)TUNEL法病理检查发现，GM3可诱导胶质瘤细胞发生凋亡。GM3给药后1~3d，大鼠脑内可检测到凋亡细胞，且仅见于胶质瘤细胞团的表面。为了探讨GM3是否对胶质瘤细胞的侵袭有干扰作用，采用Boyden小室实验和切片培养模型侵袭实验。(4)GM3抑制胶质瘤细胞的运动和侵袭，其作用依赖于GM3的浓度。局部应用GM3作为控制胶质瘤的一种新策略，可能是一种有效和安全的方法。

To investigate whether gangliosides modulate the progression of glioma cells, GM3, the simplest ganglioside, was administered into the culture of glioma cells and into the glioma-bearing rats via the cistema magna. (1) Four primary glioma cells and 4 glioma cell lines were cultured with GM3. MTT assay demonstrated that the glioma pells were inhibited those proliferation by GM3 with the concentration of 20 to 200 micromole(P<0.05). The inhibition was dependent on the concentration of GM3. (2) Rat glioma model for the in vivo analysis was made by the inoculation of C6 glioma cells into the rat meningeal space. Treated rats (n=20), those were administered GM3 on 10 days after glioma-cell-inoculation, survived 22.0 days, whereas the control rats(n=15) survived 19.5 days(P<0.05). Acute toxicity was not detected on the rats after GM3 administration. (3) On the pathological examination by TUNEL method,GM3 was found to induce apoptosis on the glioma cells. The apoptotic cells were detected from 1 to 3 days after administration of GM3 in the rat brain and detected only on the surface of the mass of glioma cells. Those results demonstrated that GM3 does not infiltrate well into the brain or into the mass of glioma cells.To investigate whether GM3 interfere with the invasion of the glioma cells, Boyden chamber assay and invasion assay with slice culture model were performed.(4) As a result, glioma cells were inhibited its motility and invasion by GM3, that was dependent on the concentration of GM3.These results demonstrate that GM3 inhibits both the proliferation and invasion of glioma cells. The local administration of GM3 as a new strategy for the control of glioma may be a useful and safe method.