摘要
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Nrg1α(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Nrg1α、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。
Objective To investigate the regulation of Neuregulin-1 （ Nrgl ） on cell adhesion molecule L1 expres- sion and its effects on the migration of U87-MG cells. Methods Cells were administered with recombinant Nrglα （rNrglct） for 24 hours （h） and 48 h, and RT-PCR was used to investigate the mRNA levels of L1. Cells were then treated with rNrglα and rNrglβ at 2.5 nmol/L for 48 h, and Western blot was applied to study the expression of L1 in response to Nrgl. siRNA targeting Nrgl was also applied to study its effects on L1 expression. In addition, cells treated with Nrgl siRNA was also subjected to wound healing assay. Immunofluorescence staining was used to study Nrglα/β and L1 expression in cells on the wound edge. Results Administration of Nrglα at 2.5 nmol/L can appartently increase the L1 mRNA levels at 24 h and 48 h time points. In addition, both 2.5 nmol/L of Nrgl α and Nrg1β can significantly increase the protein level of L1 at 48 h （P 〈 0. 01 and P 〈 0. 05）. siRNA targeting Nrgl for 48 h can appartently reduce the expression of Nrgl, accompanied with the reduction of L1 levels. Wound healing assay indicated that downregulating Nrgl 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 Nrgl can regulate L1 expression in human glioblastoma U87-MG cells, which may partially contrib- ute to cell migration.
出处
《基础医学与临床》
CSCD
北大核心
2013年第7期829-833,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(81171138)
