人脐静脉内皮细胞损伤后迁移能力的变化与血管内皮钙黏蛋白和连环蛋白p120的关系

Change of Migration Ability of Human Umbilical Vein Endothelial Cell After Injury and Its Relationship with Vascular Endothelial Cadherin and p120 Catenin

目的初步探讨人脐静脉内皮细胞(HUVEC)损伤之后迁移能力的变化与血管内皮钙黏蛋白(VE-cad)和连环蛋白p120(p120ctn)的关系。方法 DMEM培养基培养HUVEC,将HUVEC分为对照组和损伤组。Transwell实验检测HUVEC迁移能力的变化。Western blot测定p120ctn与VE-cad蛋白表达水平。免疫荧光实验检测VEcad的定位表达变化。免疫共沉淀法检测p120ctn与VE-cad的相互结合。结果 Transwell实验发现HUVEC经损伤刺激6、8 h后迁移能力最强(P<0.05)。Western blot结果显示HUVEC损伤6、8 h后p120ctn及VE-cad表达水平明显上调。免疫荧光实验显示HUVEC经损伤刺激后,VE-cad的定位由细胞膜转到细胞浆。免疫共沉淀证实p120ctn可以与VE-cad相互结合。结论 HUVEC损伤刺激后迁移能力增强,其机制可能与升高的p120ctn将VEcad由细胞膜携带入细胞浆导致VE-cad膜表达缺失有关。

Aim To explore preliminarily the change of migration ability of human umbilical vein endothelial cell（ HUVEC） after injury and its relationship with vascular endothelial cadherin（ VE-cad） and p120 catenin（ p120ctn）.Methods HUVEC was cultured in MEM culture medium,and was divided into control group and injury group. The change of HUVEC migration ability was detected by Transwell experiment. Expression levels of P120 ctn and VE-cad protein were determined by Western blot. Location expression change of VE-cad was detected by immunofluorescence assay.Mutual combination of P120 ctn and VE-cad was detected by Co-immunoprecipitation. Results Transwell experiment showed that the migration ability of HUVEC was the strongest in the 6 and 8 h after injury（ P〈0. 05）. Western blot results showed that p120 ctn and VE-cad expression levels of HUVEC were significantly up-regulated in the 6 and 8 h after injury. Immunofluorescence assay showed that VE-cad was localized from cell membrane into cytoplasm after HUVEC was injured. Co-immunoprecipitation confirmed that P120 ctn could be mutually combined with VE-cad. Conclusion The migration ability of HUVEC is enhanced after injury stimulation,and its mechanism may be related to that elevated p120 ctn carries VE-cad from cell membrane into cytoplasm,which leads to loss of VE-cad expression on the membrane.