神经调节因子Neuregulin-1(Nrg1)调节U87-MG细胞的黏附分子L1表达及促迁移作用

摘要

Objective To investigate the regulation of Neuregulin-1 (Nrg1) on cell adhesion molecule L1 expression and its effects on the migration of U87-MG cells.Methods Cells were administered with recombinant Nrg1 α(rNrg1ot) for 24 hours (h) and 48 h,and RT-PCR was used to investigate the mRNA levels of L1.Cells were then treated with rNrg1 α and rNrg1 β at 2.5 nmol/L for 48 h,and Western blot was applied to study the expression of L1 in response to Nrg1.siRNA targeting Nrg1 was also applied to study its effects on L1 expression.In addition,cells treated with Nrg1 siRNA was also subjected to wound healing assay.Immunofluorescence staining was used to study Nrg1α/β and L1 expression in cells on the wound edge.Results Administration of Nrg1α at 2.5 nmol/Lcan appartently increase the L1 mRNA levels at 24 h and 48 h time points.In addition,both 2.5 nmol/L of Nrg1 αand Nrg1 β can significantly increase the protein level of L1 at 48 h (P ＜ 0.01 and P ＜ 0.05).siRNA targeting Nrg1 for 48 h can appartently reduce the expression of Nrg1,accompanied with the reduction of L1 levels.Wound healing assay indicated that downregulating Nrg1 expression not only reduced the expression of L1 in cells on the wound edge,but also reduced cell migration,as was indicated by the wound space with a higher width.Conclusions Nrg1 can regulate L1 expression in human glioblastoma U87-MG cells,which may partially contribute to cell migration.%目的 研究神经调节因子Neuregulin-1(Nrg1)对人胶质母细胞瘤U87-MG细胞中细胞黏附分子L1表达及细胞迁移的影响.方法 给予U87-MG细胞重组Nrg1d(rNrg1α,2.5 nmol/L)24和48 h,RT-PCR观察L1 mRNA表达；给予细胞rNrg1α或rNrg1β(2.5 nmol/L)48 h,Western blot检测L1蛋白水平.用Nrg1 siRNA处理细胞,Western blot观察L1蛋白水平,用细胞划伤实验观察细胞迁移.结果 Nrg1α可促进L1 mRNA表达；与0 nmol/L组相比,Nrg1α及Nrg1β(2.5 nmol/L)均可显著增加L1蛋白水平(P＜0.01,P＜0.05).与对照siRNA相比,Nrg1 siRNA明显降低Nrg1表达,并伴有L1表达下降.Nrg1 siRNA处理细胞划伤16 h,划伤边缘细胞Nrg1 α、Nrg1β及L1荧光信号降低,细胞迁移减弱.结论 Nrg1可调节U87-MG细胞L1表达,其参与细胞迁移可能与提高L1表达有关.