甲基-β-环糊精干扰脂质微区影响脑胶质瘤细胞系C6侵袭和迁移的体外研究

Study on the effects of membrane lipid microdomain extractor methyl-β-cyclodextrin on invasion and migration of cerebral glioma cell line C6 in vitro

目的观察甲基-β-环糊精(MβCD)破坏脂质微区结构对经体外培养的大鼠脑胶质瘤细胞系C6细胞侵袭和迁移能力的影响。方法采用MβCD(5mmol/L)破坏经体外培养的大鼠脑胶质瘤细胞系C6细胞膜脂质微区,通过Transwell细胞侵袭和迁移实验检测细胞侵袭性和迁移能力;Western blotting检测细胞膜脂质微区结构蛋白Caveolin-1和膜受体表皮生长因子受体表达水平的变化。结果干扰组大鼠脑胶质瘤细胞系C6细胞Caveolin-1和表皮生长因子受体表达水平明显低于对照组(P=0.000,0.001)。对照组大鼠脑胶质瘤细胞系C6细胞穿过聚碳酸酯微膜数目为(54.00±9.59)个/视野,干扰组为(23.94±5.56)个/视野,两组比较差异有统计学意义(P=0.000)。对照组大鼠脑胶质瘤细胞系C6细胞迁移至空白端的细胞数目为(134.00±9.68)个/视野,干扰组为(88.00±6.22)个/视野,两组比较差异有统计学意义(P=0.000)。结论甲基-β-环糊精破坏脂质微区结构后可降低经体外培养的大鼠脑胶质瘤细胞系C6细胞的侵袭性和迁移能力,其机制可能与降低Caveolin-1和表皮生长因子受体等多种膜蛋白和受体表达水平有关。

Objective To study the effects of the destruction of membrane lipid microdomain with extractor methyl-β-cyclodextrin （MβCD） on invasion and migration of cerebral glioma cell line C6 in vitro. Methods The membrane lipid microdomain of cerebral glioma cell line C6 in vitro were destructed with MβCD （5 mmol/L）. Abilities of cell invasion and migration were examined by Transwell and ＂Wound-healing＂ method, respectively. Expressions of Caveolin-1, the structural protein of membrane lipid microdomain, and epidermal growth factor receptor （EGFR）, one of the membrane receptors were measured by Western blotting. Results In comparison with control group, expressions of Caveolin-1 and EGFR of cerebral glioma cell line C6 were significantly decreased in interference group, the differences were significant （P=0.000, 0.001）. The number of cells invasion through the Matrigel basement membrane for cerebral glioma cell line C6 in control group was （54.00 ± 9.59）/visual field, and it was （23.94 ± 5.56）/visual field in interference group. The difference between 2 groups was significant （P=0.000）. The number of cells migration into the margin for cerebral glioma cell line C6 in control group was （134.00 ± 9.68） / visual field, and it was （88.00 ± 6.22） / visual field in interference group. Significant difference was also seen between 2 groups （P=0.000）. Conclusion Membrane lipid microdomain destruction by the MI3CD can decrease the invasion and migration of cerebral glioma cell line C6 in vitro. It maybe related to the decrease of expressions of the membrane protein and receptors such as Caveolin-1 and EGFR.