神经细胞黏附分子对小鼠骨髓间充质干细胞黏附、迁移及细胞形态的影响

Effect of neural cell adhesion molecule on adhesion,migration and morphology of mouse BMSCs

目的探讨神经细胞黏附分子(NCAM)对小鼠骨髓间充质干细胞(BMSCs)黏附、迁移及细胞形态的影响。方法从野生型小鼠和NCAM基因敲除小鼠中分离、培养BMSCs,Western blot和免疫荧光标记检测NCAM的表达;划痕实验和黏附实验分别检测细胞迁移和黏附能力;倒置显微镜下观察细胞形态;Western blot检测β1整联蛋白、E-cadherin、β-catenin和N-cadherin的表达。结果 NCAM基因敲除后细胞的迁移能力和黏附能力明显降低,β1整联蛋白的表达下调(P<0.01),BMSCs形态由不规则形变为扁平形,且成簇增殖,上皮细胞标记蛋白E-cadherin和β-catenin的表达显著上调(P<0.05),而间质细胞标记蛋白N-cadherin的表达下调(P<0.01)。结论NCAM通过调控β1整联蛋白的表达影响BMSCs的黏附和迁移,同时NCAM可能对BMSCs的间质上皮转化起负调控作用。

Objective To explore the effect of neural cell adhesion molecule （NCAM） on adhesion, migration and morphology of mouse bone marrow-derived mesenchymal stem cells （BMSCs） .Methods We isolated and cultured BMSCs from wild-type and NCAM gene knockout mice. The expression of NCAM was detected by Western blot and immunofluorescence. Wound healing and adhesion assays were used to detect cell migration and adhesion ability re- spectively. The morphological changes were observed and the expressings of protein β1 integrin, E-cadherin, β-eatenin and N-cadherin were analysed by Western blot. Results The migration and adhesion of BMSCs were significantly reduced after NCAM gene knockout. Meanwhile, the expression of β1 integrin was lower than those in wild-type BMSCs （P〈0.01）. The morphology of NCAM gene knockout BMSCs changed from irregular to flat- tened, and expressed epithelial identification marker E-cadherin and β-catenin （P〈0.05）. However, the ex- pression level of mesenchymal identification marker N-cadherin was decreased （P〈0.01）. Conclusions NCAM is involved in adhesion and migration of BMSCs via regulating the expression of β1 integrin. Furthermore,NCAMmay negatively regulate the mesenchymal-epithelial transitions of BMSCs.