Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppress...Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppression VEs and VEs transfected with CREG gene modified adenovirus(Ad-CREG) were cultured with dulbecco’s modified eagle’s medium contained 10%fetal calf serum. Western blot was used to detect the protein level of CREG and integrin-linked kinase(ILK) in the four kind ECs.Tran-swell migration model was applied to compare the migration cell number of the four kind ECs.Two kinds of ILK mutant plasmids;PCXN2-flag-ILK wt-IRES-GFP(wild-type ILK)and PCXN2-flag-ILK p-parvin-IRES-GFP(P-parvin-binding mutant) were used to transfect VS and VE respectively,then the two kind transfection ECs were named as VS-wtILK and VE-P -parvin which were selected by G418(600ng/ml)for 2 weeks;Transwell migration model was applied to compare migration capability before and after ILK plasmids transfecting VE and VS.Results Western blot analysis showed that CREG overexpression promoted ILK expression in ECs,on the contrary,ILK expression was down-regulated in CREG silent ECs(P【0.05).Further more,ILK expression was up-regulated obviously in VE transfected with Ad-CREG(P【 0.05);Transwell migration model showed that EC’s migration capability was positively correlated with the expression level of CREG in EC,that is,CREG overexpression induced VE migration and CREG silent suppressed VE migration, moreover,Ad-CREG transfecting VE showed better migration capability accompanied with CREG expression increase by transwell migration model(P【0.05).In order to know the relationship between ILK expression and cell migration,we obtained stable transfection cell strains of VS-wtILK and VE-Pparvin, transwell migration model demonstrated that VS-wtILK remarkably corrected the poor migration capability of VS(P【 0.01),butβ-parvin combining site mutation in ILK genes inhibited VE migration markedly(P【0.01).Conclusions ILKp -parvin signal pathway mediated vascular endothelial cell migration induced by CREG.展开更多
文摘Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppression VEs and VEs transfected with CREG gene modified adenovirus(Ad-CREG) were cultured with dulbecco’s modified eagle’s medium contained 10%fetal calf serum. Western blot was used to detect the protein level of CREG and integrin-linked kinase(ILK) in the four kind ECs.Tran-swell migration model was applied to compare the migration cell number of the four kind ECs.Two kinds of ILK mutant plasmids;PCXN2-flag-ILK wt-IRES-GFP(wild-type ILK)and PCXN2-flag-ILK p-parvin-IRES-GFP(P-parvin-binding mutant) were used to transfect VS and VE respectively,then the two kind transfection ECs were named as VS-wtILK and VE-P -parvin which were selected by G418(600ng/ml)for 2 weeks;Transwell migration model was applied to compare migration capability before and after ILK plasmids transfecting VE and VS.Results Western blot analysis showed that CREG overexpression promoted ILK expression in ECs,on the contrary,ILK expression was down-regulated in CREG silent ECs(P【0.05).Further more,ILK expression was up-regulated obviously in VE transfected with Ad-CREG(P【 0.05);Transwell migration model showed that EC’s migration capability was positively correlated with the expression level of CREG in EC,that is,CREG overexpression induced VE migration and CREG silent suppressed VE migration, moreover,Ad-CREG transfecting VE showed better migration capability accompanied with CREG expression increase by transwell migration model(P【0.05).In order to know the relationship between ILK expression and cell migration,we obtained stable transfection cell strains of VS-wtILK and VE-Pparvin, transwell migration model demonstrated that VS-wtILK remarkably corrected the poor migration capability of VS(P【 0.01),butβ-parvin combining site mutation in ILK genes inhibited VE migration markedly(P【0.01).Conclusions ILKp -parvin signal pathway mediated vascular endothelial cell migration induced by CREG.
