神经酰胺信号通路在花生四烯酸乙醇胺抑制人U251胶质瘤细胞侵袭中的作用

Effect of ceramide signaling pathway on anandamide-mediated inhibition of adhesion and invasion of U251 glioma cell line

目的 观察神经酰胺信号通路在花生四烯酸乙醇胺（AEA）抑制人胶质瘤U251细胞黏附和侵袭生物学行为的抑制作用.方法 通过5 μmol/L AEA、5μmol/L AEA＋ 10 μmol/L C2-神经酰胺作用24h的预处理U251细胞,同时与先用10 μmol/L烟曲霉毒素（FB1）预处理U251细胞24 h后的实验组比较,应用噻唑蓝（MTT）比色法测定U251细胞黏附作用的影响,应用Transwell小室实验检测U251细胞侵袭能力的影响.结果 5μmol/L AEA、5μmol/L AEA＋ 10 μmol/L C2-神经酰胺均能抑制U251细胞侵袭生物学行为的作用,5μmol/L AEA组U251细胞黏附和侵袭能力分别降低66.53％和83.85％,5μmol/L AEA＋ 10 μmol/L C2-神经酰胺组分别降低71.64％和86.43％;而用10 μmol/L FB1先预处理U251细胞24 h后,5μmol/L AEA组U251细胞黏附和侵袭能力仅分别降低27.43％和32.23％,5μmol/L AEA＋ 10 μmol/L C2-神经酰胺组则分别降低56.61％和68.11％.结论 AEA具有人胶质瘤U251细胞黏附和侵袭生物学行为的抑制作用,这种作用可能是通过慢反应神经酰胺信号通路来实现的.

Objective To observe the effect of ceramide signaling pathway on anandamide （AEA）-mediated inhibition of adhesion and invasion of U251 glioma cell line.Methods Human U251 cells were cultured with DMEM in vitro.Cultured cells were divided into control group,5 μmol/L AEA group,and 5 μmol/L AEA ＋ 10 μmol/L C2-ceramide group.At the same time,the AEA groups were pretreated with 10 μmol/L fumonisin B1 （FB1） for 24 h early.The cell adhesion rate of U251 cells was examined by methyl thiazol tetrazolium （MTT） assay at different time points of AEA treatment.The abilities of invasion were detected by Transwell assay.Results 5 μmol/L AEA,and 5 μmol/L AEA ＋ 10 μmol/L C2-ceramide could inhibit invasive abilities of U251 cells.The capabilities of adhesion and invasion of U251 cells in 5 μmol/L AEA group was reduced by 66.53％ and 83.85％ respectively （P ＜ 0.01）,and those in 5 μmol/L AEA ＋ 10 μmol/L C2-ceramide group were decreased by 71.64％ and 86.43％ respectively （P ＜0.01）.After pretreatment of U251 cells with 10 μmol/L FB1 for 24 h,The capabilities of adhesion and invasion of U251 cells in 5 μ mol/L AEA group were only decreased by 27.43％ and 32.23％,and those in 5 μmol/L AEA ＋ 10 μmol/L C2-ceramide group decreased by 56.61％ and 68.11％ （P ＜0.01）,respectively.Conclusion AEA can inhibit the capabilities of adhesion and invasion of U251 cells via the ceramide signaling de novo synthesis pathway.