大鼠小胶质细胞及巨噬细胞对胶质瘤C6细胞株迁移能力的影响

目的 探讨大鼠小胶质细胞及外周血巨噬细胞对胶质瘤C6细胞株迁移能力的影响.方法 将大鼠小胶质细胞及外周血单核细胞诱导分化形成的巨噬细胞分别与胶质瘤C6细胞共培养,以大鼠成纤维细胞共培养为阳性对照,单纯胶质瘤C6细胞为空白对照,共培养0、24、48 h分别行划痕实验,于划痕后6 h观察划痕实验结果;共培养24、48 h时以CD11b标记、流式分选分离小胶质/巨噬细胞及C6细胞,行Western blot检测0、24、48 h小胶质/巨噬细胞IL-10、IL-18和MMP-9及C6细胞株TGF-β、FAS配体表达水平的改变.结果 早期小胶质细胞及巨噬细胞均抑制胶质瘤的迁移,巨噬细胞的抑制效果更强(P<0.05);与胶质瘤细胞共培养后两种细胞对肿瘤的抑制作用消失且差异无统计学意义,均表现为促进肿瘤迁移.小胶质细胞、巨噬细胞在初始阶段IL-10、IL-18、金属基质蛋白酶MMP-9表达略有不同,但共培养后两种细胞的上述蛋白表达均升高且一致;共培养后的胶质瘤细胞的TGF-β、FAS配体较共培养前表达均升高且一致.结论 小胶质细胞及巨噬细胞与C6胶质瘤细胞共培养后被驯化成同样的GAMs,对C6细胞迁移影响的生物学表现一致.

Effects of microglia and macrophages on migration of rat C6 glioma cells

Objective To determine the effect of microglia and peripheral blood macrophages on the migration of C6 glioma cells.Methods The rat microglial cells and macrophages differentiated from peripheral blood mononuclear cells were co-cultured with C6 glioma cells.Co-cultured with rat fibroblasts served as positive control and singly cultured C6 cells as blank control.Scratch wound healing assay was carried in 0, 24 and 48 h after co-culture, and the results were observed in 6 h later.In 24 h and 48 h after co-culture, CD11b+ flow cytometry was used to sort and isolate microglia/macrophages and C6 cells.The expression levels of IL-10, IL-18 and MMP-9 in microglia and macrophages and those of TGF-β and FAS ligand in glioma cells were detected by Western blotting when co-cultured for 0, 24 and 48 h.Results At the initial state, both microglia and macrophages inhibited the migration of glioma, and the inhibitory effect of macrophage was stronger (P<0.05).The tumor inhibitory effect of 2 kinds of cells disappeared after co-culture with glioma C6 cells, and all of them showed the effect of promoting tumor migration.The expression levels of IL-10, IL-18 and MMP-9 in the microglia and macrophages were different in the initial stage.But the levels of these proteins were increased in these cells and consistent after co-culture.The expression levels of TGF-β and FAS ligand were also increased and consistent in glioma cells C6 after co-culture.Conclusion Microglia and macrophages are domesticated into the same glioma associated microglia/macrophages after co-culture with C6, and 2 kinds of cells show the same biological effects when co-culture with C6 cells.