Effects of downregulation of astrocyte up-regulated gene-1 expression combined with all-trans retinoic acid on vascular mimicry in glioma
Zhao Wanying
Zhang Binbin
Li Ruichun
Liang Chen
Abstract：Objective To investigate the effects and potential mechanisms of downregulation of astrocyte up-regulated gene-1(AUG-1)expression combined with all-trans retinoic acid(ATRA)on vasculogenic mimicry(VM)formation in glioma.Methods U87 glioma cells were transfected with AEG-1 shRNA lentiviral vectors(U87-siAEG-1)and incubated in a medium containing 20µmol/L ATRA.A Matrigel-based tube formation assay was performed to evaluate VM formation in vitro.They were divided into control group(U87 cells),ATRA group(U87 cells + 20μmol/L ATRA),siCon group(U87-siCon cells),siCon+ATRA group(U87-siCon cells+20μmol/L ATRA),siAEG-1 group(U87-siAEG-1 cells)and siAEG-1+ATRA group(U87-siAEG-1 cells + 20μmol/L ATRA).In vitro lumen formation assay was performed for evaluating in vitro vascular mimicry formation ability of U87 cells.Real-time PCR and Western-blot analysis were conducted to investigate the expression of related gene.Glioma xenograft models were generated via subcutaneous implantation of glioma cells in nude mice.Tumor-bearing mice received an intraperitoneal injection of 10mg/kg ATRA.It was divided into:control group(U87 cell modeling),ATRA group(U87 cell modeling + ATRA intervention),siCon group(U87-siCon cell modeling),siCon + ATRA group(U87-siCon cell modeling + ATRA intervention),siAEG-1 group(U87-siAEG-1 cell modeling)and siAEG-1+ATRA group(U87-siAEG-1 cell modeling + ATRA intervention).CD34/periodic acid-Schiff double staining was performed to detect VM channels in vivo.The size of tumors was measured,and the tumor growth curve was drawn to evaluate the tumor growth.Results A combination of ATRA intervention and downregulation of AEG-1 expression(siAEG-1+ATRA group)significantly decreased VM formation of glioma in vitro and in vivo,and significantly inhibit the growth of subcutaneous glioma xenografts.The expression levels of matrix metalloproteinase(MMP)-2,MMP-9 and VE-cadherin in glioma cells significantly decreased after intervention.Compared with the control group,the difference was statistically significant(all P<0.05).Conclusions Downregulation of AEG-1 combined with ATRA can significantly inhibit the formation of VM and the growth of glioma.The mechanism may be related to the inhibition of the expression of MMP-2,MMP-9 and VE-cadherin.
Keywords：gliomaastrocyte elevated gene-1all-trans retinoic acidvasculogenic mimicry
Publication Date：2024-01-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 24-29 )