The Research Of CELSR1Promotes Endothelial Cell Migration And The Formation Of Vessels

Objective：Using GWAS analysis, Yoshiji Yamada group identified CELSR1as asusceptibility gene for ischemic stroke（IS） in Japanese individuals. Our grouppreliminary experiments also identified CELSR1gene single nucleotidepolymorphism loci rs6007897and rs4044210was significantly associated withischemic stroke, associated with aortic atherosclerosis。CELSR1is a key componentof the non-canonical Wnt/PCP pathway, and the non-canonical Wnt/PCP pathway wasrecently found to play a critical role in angiogenesis. We transfected CELSR1shRNAand over expression CELSR1to mute and overexpression the CELSR1in humanaortic endothelial cell（HAEC） by liposomes instant transfection and Lentiviralvector to explore the effects of CELSR1expression on HAECs and aorticatherosclerosis.Methods：Constructed a plasmid with shCELSR1and input it into HAEC cell withlentivirus. CELSR1sequence was transfected into HAEC by TALEA technology,and cell lines was selected by puromycin. The expression level of CELSR1wasdetermined by Western blot and Real time-PCR, the cell proliferation and migrationwas detected by MTT assay,cyclization and cell scratch.Results: ShCELSR1sequence can inhibit the expression of CELSR1effectively, andthe mRNA relative expression level was improved by transfected the CELSR1sequences, compared with the control group,and the difference was statisticallysignificant (P <0.05). MTT assay results show that the Proliferation of HAEC wasinhibited by interference CELSR1compared with control cells, the difference wasstatistically significant (P <0.05); The Proliferation of HAEC was promoted byoverexpression CELSR1,the difference was statistically significant (P <0.05);Transwell experiments and cell scratch showed that the ability of migration ofoverexpressing CELSR1was stronger than the control group, and the difference wasstatistically significant (P <0.05); The ability of migration of CELSR1shRNA wasweaker than the control group, and the difference was statistically significant (P <0. 05); Ring formation assay showed that the ring numbers formed by overexpressingCELSR1was much more the control group, and the difference was statisticallysignificant (P <0.05). Ring formation assay showed that the ring numbers formed byCELSR1shRNA was much little the control group, and the difference wasstatistically significant (P <0.05).Conclusion: CELSR1shRNA could knock down the expression of CELSR1inHAECs, and can inhibit the proliferation,migration,cyclization of HAEC effectively.Transfection CELSR1could increase the expression of endogenous CELSR1ofHAEC, and can promote the proliferation and migration, cyclization of HAECeffectively. The result suggest that CELSR1may play an important role in the processof angiogenesis.