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表皮生长因子诱导人脐静脉内皮细胞迁移过程中波形蛋白及基质金属蛋白酶2的表达变化
卢诗军 杨慧科 马文平 李晓冬 张雅芳*
解剖学报 ›› 2013, Vol. 44 ›› Issue (6) : 789-794.
表皮生长因子诱导人脐静脉内皮细胞迁移过程中波形蛋白及基质金属蛋白酶2的表达变化
Changes of vimentin and matrix metalloproteinases-2 expressions in epidermal grouth factor induced migration of human umbilical vein endothelial cells
目的 观察人脐静脉内皮细胞（HUVECs）迁移过程中表皮生长因子（EGF）对波形蛋白（vimentin）和基质金属蛋白酶2（MMP-2）表达的影响，分析波形蛋白与EGF促HUVECs迁移能力的关系。 方法 EGF处理HUVECs细胞10h后，采用细胞划痕实验检测HUVECs细胞的迁移水平，荧光显微镜下观察细胞的形态变化情况；免疫荧光染色观察划痕诱导HUVECs迁移时波形蛋白的表达变化；Western blotting和RT-PCR法分析MMP-2及波形蛋白表达。 结果 EGF刺激后HUVECs的迁移能力上升1.8倍，迁移细胞的形态发生改变，由铺路石样变为长梭形，同时波形蛋白和MMP-2的表达上调（P<0.05）。 结论 EGF促进HUVECs的迁移过程中可明显上调波形蛋白和MMP-2的表达，波形蛋白与HUVECs的迁移相关。
Objective To evaluate the relationship between vimentin and migration induced by epidermal growth factor (EGF) in human umbilical vein endothelial cells (HUVECs), the expressions of vimentin and matrix metalloproteinases-2 (MMP-2) were examined in epidermal growth factor (EGF) -induced migration of HUVECs. Methods In the current study, the migration of HUVECs was analyzed after EGF stimulation for 10h by scratch assay, and the effect of EGF on the cells morphology was also observed. The monolayer HUVECs were scratched to induce the cells migration in both control group and EGF group, and then immunofluorescent staining was used to detect the vimentin expression. Both vimentin and MMP-2 expressions were assessed via Western blotting and RT-PCR. Results The scratch assay revealed a 1.8-fold increase induced by EGF in migration levels of HUVECs. Interestingly, the HUVECs morphology also altered dramatically from cobblestone-like shape to spindle-like shape after EGF stimulation. As a result of EGF stimulation, both vimentin and MMP-2 expressions in HUVECs were up regulated in both protein and mRNA levels (P<0.05). Conclusion EGF upregulates both vimentin and MMP-2 expressions effectively in EGF-induced migration of HUVECs, and vimentin is closely associated with the migration in HUVECs.
表皮生长因子 / 细胞迁移 / 波形蛋白 / 基质金属蛋白酶2 / 免疫印迹法
Epidermal growth factor / Migration / Vimentin / Matrix metalloproteinase 2 / Western blotting
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