Si-WDR4 encapsulated by the brain-targeting peptide Angiopep2 inhibits glioblastoma proliferation via phosphatidylinositol 3 kinase/protein kinase B signaling
Objective To investigate the effect of brain-targeted peptide,Angiopep2 encapsulated with si-WDR4 on glioblastoma and its potential mechanism.Methods Angiopep2/si-RNA was prepared,and its encapsulation efficiency and uptake rate were detected.In vitro,U87 cells were treated with phos-phate buffer saline(PBS),Angiopep2/si-NC,si-WDR4 and Angiopep2/si-WDR4,respectively.The mRNA expression level of WDR4 was detected by RT-qPCR,the protein expression levels of WDR4,phos-phatidylinositol 3 kinase(PI3K),and protein kinase B(Akt)were detected by Western blotting,the via-bility of U87 cells was detected by cell counting kit-8(CCK-8)assay,and the proliferation of U87 cells was detected by 5-Ethynyl-2'-deoxyuridine(EdU)staining.In vivo,a xenograft tumor model was estab-lished by U87-Luc cells.After 10 days,PBS,Angiopep2/si-NC,si-WDR4,and Angiopep2/si-WDR4 were injected through the tail vein,respectively.On the 28th day,the dual-luciferase activity of the tumor was detected by small animal in vivo imaging,and the tumor size was detected by hematoxylin and eosin(HE)staining.The quantitative data were compared using t test.Results Angiopep2 had a good binding ability to si-WDR4,forming a complete complex at an N/P ratio of 3∶1,and could promote the uptake of si-RNA by tumor cells;in vitro,compared with Angiopep2/si-NC,Angiopep2/si-WDR4 treatment could reduce the mRNA(0.21±0.14,t=4.13,P＜0.05)and protein levels(0.42±0.17,t=3.53,P＜0.05)of WDR4 in U87 cells,reduce the expression of PI3K(0.34±0.19,t=5.17,P＜0.05)and Akt(0.24±0.17,4.26,P＜0.05),decrease the activity of U87 cells(52.00±11.33,t=4.59,P＜0.05),and decline the number of EdU-positive cells(23.00±7.28,t=12.12,P＜0.05);in vivo,the dual luciferase activity in the brain tissue of nude mice treated with Angiopep2/si-WDR4 was declined(10.02±2.06,t=6.14,P＜0.05).The relative volume of tumors was reduced(0.38±0.17,t=7.42,P＜0.05).Conclusion Angiopep2/si-WDR4 can promote the uptake of si-WDR4 by tumor cells and in-hibit the proliferation of glioblastoma.
GlioblastomaAngiopep2/si-WDR4Cell proliferationPhosphatidylinositol 3 kinase/protein kinase B signaling pathway
NETL
NSTL
万方数据
