AMD3100 Inhibits The Migration And Differentiation Of Neural Stem Cells By Down-regulation Of CXCR4/?-catenin In Vitro

Objective:To investigate the inhibition effect of AMD3100 on the proliferation,migration and differentiation of central neural stem cells by CXCR4/?-catenin signaling pathway.Methods:1.Preparation of AMD3100 containing different concentrations(0ug/L,40ug/L,80ug/L,160ug/L,320ug/L,640ug/L and 1000ug/L)in the culture medium.Central neural stem cells was cultured by suspension method.The OD value of each group was detected by CCK8 method at 2 hours,24 hours,48 hours and 72 hours respectively,and the proliferation inhibition rate of each group was calculated at different times.2.The drug concentration and reaction time(80ug/L,48h)were selected from the previous experiment.The control group(0ug/L)and AMD3100 group(80ug/L)cells were cultured for 48 hours.Wound Healing was used to detect the migration ability of central neural stem cells and Western Blot was used to detect the expression of Nestin,?-tubulin,Wnt,?-catenin and ?-catenin(P-Tyr489).Results:1.CCK8 assay results: With the increase of the extension time,in 80ug/L,160ug/L,320ug/L,640ug/L,and 1000ug/L,the proliferation capacity was gradually decreased and the inhibition rate was gradually increased.At 48 hours and 72 hours,the proliferation capacity was significantly lower than that of 0 ug/L(P <0.01),and the inhibition rate of all was more than 50%.The cell proliferation ability of 40ug/L was no significant difference compared with 0ug/L(P> 0.05).At 2 hours and 24 hours,the inhibition rate was gradually increased with increasing concentration.At 48 hours and 72 hours,the inhibition rates of 160ug/L,320ug/L and 640ug/L with a increasing trend were lower than 80ug/L and 1000ug/L,respectively,but higher than all concentrations of 24 hours.2.The Wood Healing results: At 48 hours,percentage of the area of cell migration in AMD3100 group(19.80% ± 1.83)was significantly lower than the control group(32.92% ± 2.64),the difference was statistically significant(P<0.05).Western Blot results: The expression of Nestin,?-tubulin,?-catenin and ?-catenin(P-Tyr489)in AMD3100 group was lower than that in control group(P<0.05).There was no significant difference in Wnt expression between the two groups(P> 0.05).Conclusion:1.AMD3100 inhibit the proliferation of neural stem cells in a time and concentration-dependent.2.AMD3100 could inhibit the proliferation,migration and differentiation of neural stem cells.3.AMD3100 may be through CXCR4/?-catenin pathway to inhibit the proliferation and migration and differentiation of neural stem cells.