Effect of aloe-emodin combined with targeted inhibition of FOXC1 gene on the biological behavior of glioma cells
Objective To investigate the effects of aloe-emodin(AE)combined with siRNA-FOXC1 targeted silencing of the FOXC1 gene on regulating the Wnt/β-catenin pathway on the proliferation,apoptosis,and migration of glioma U251 cells.Methods The study was conducted at the Medical Experimental Center of Binzhou Medical University Hospital from June 2022 to December 2023.The siRNA sequence(FOXC1-siRNA-2102)with the highest efficiency in targeting silencing of FOXC1 gene was selected,and the cells were transfected by liposome method.MTT method was used to detect the effects of different concentrations of AE on the proliferation of glioma U251 cells,and the IC50 value was calculated.CCK8 method was used to detect the changes of cell proliferation ability at 24 h,48 h,72 h,and 96 h after transfection.Cells were divided into a control group,a AE group,a siRNA-FOXC1 group,and a siRNA-FOXC1+AE group,and the effect on apoptosis in each group was detected by Annexin V-PE/7-AAD staining-flow cytometry.Transwell assay was used to detect the migration ability of cells in each group,qPCR assay was used to detect the mRNA expression level of the FOXC1 gene in each group,and Western blotting method was used to detect the protein expression levels of FOXC1,β-catenin,and C-myc in each group.F-test was used for statistical analysis.Results With the increase of AE dose,the proliferation inhibition rate of glioma U251 cells was significantly increased[(0.04±12.86)％in the control group,(24.45±6.96)％in the 30 μmol/L group,(48.41±5.67)％in the 60 μmol/L group,(60.63±9.96)％in the 90 μmol/L group,and(78.23±13.34)％in the 120 μmol/L group],and the IC50 of AE in U251 cells was 61.43 μmol/L.The cell proliferation of the siRNA-FOXC1 group,AE group,and siRNA-FOXC1+AE group was higher than that of the control group over time,and the proliferation of the siRNA-FOXC1+AE group was the most significantly decreased;the apoptosis rates of the siRNA-FOXC1 group,AE group,and siRNA-FOXC1+AE group were higher than that of the control group over time,and the cell proliferation of siRNA-FOXC1+AE group was the most obvious;the mRNA expression levels of FOXC1 and protein expression levels of β-catenin,C-myc,and FOXC1 of the siRNA-FOXC1 group,AE group,and siRNA-FOXC1+AE group were lower than those of the control group over time,and those of the siRNA-FOXC1+AE group were the most significantly decreased(all P＜0.05).The cell migration abilities of the siRNA-FOXC1 group,AE group,and siRNA-FOXC1+AE group were lower than that of the control group over time,and that of the siRNA-FOXC1+AE group was the most significantly decreased[the number of migrating cells was(193.00±14.17)in the control group was,(84.00±14.22)in the AE group,(80.67±5.69)in the siRNA-FOXC1 group,and(36.33±11.59)in the siRNA-FOXC1+AE group,with a statistically significant difference(F=93.55,P＜0.05)].Conclusions AE can inhibit the proliferation and migration of glioma U251 cells and promote cell apoptosis.This mechanism is associated with FOXC1 gene regulation through Wnt/β-catenin pathway.
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