Figure 4.

Overexpression of miR-326 combined with curcumin treatment decreased GLI1 expression. (A) Real-time PCR analysis of GLI1 expression in U87 and U251 cells following miR-326 transfection and curcumin treatment for 24 h. (B) U87 and U251 cells were transfected with miR-326 or miR-Scr followed by cotransfection of a firefly luciferase reporter construct containing 8 consecutive consensus GLI1-binding sites (8×-GLI). Cells were then treated as indicated with solvent DMSO (control) or curcumin. Both firefly and Renilla luciferase activities were quantified using the Dual-Luciferase Reporter Assay System and normalized with Renilla luciferase activity. The data represent the mean ± SEM of 3 replicates (*P < 0.05; **P < 0.01, and ***P < 0.001). (C) Western blot assay showing protein GLI1 expression in glioma cells treated with miR-326, curcumin, and their combination.