Effects of short hairpin RNA targeting FOXP3 gene on proliferation and apoptosis of glioma cells

Abstract

Objective To investigate the effects of short hairpin RNA (shRNA) targeting Forkhead box P3 (FOXP3) gene on the proliferation and apoptosis of human glioma cells U87 and LN229. Methods 4 plasmids containing different sequences of shRNA targeting FOXP3 gene and 1 plasmids containing random sequences were transfected into human glioma U87 cells and LN229 cells mediated by Lipofectamine 2000. The expression of FOXP3 protein was detected by Western blot to screen the most effective shRNA plasmid. The cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry, respectively. Results At 72 h after transfection, the expression of FOXP3 protein was most obviously suppressed by shRNA-1 sequence compared with other shRNA sequences. The CCK-8 assay results showed that the proliferation rates of U87 cell and LN229 cell tranfected with shRNA-1 plasmid were (122.00±4.32)% and (118.36±2.49)% at 72 h, respectively, which were significantly higher than that transfected with neg-shRNA plasmid (neg-shRNA group) and the non-transfection group (control group) (P<0.05). At 72 h after transfection, the apoptosis rates of U87 cell and LN229 cell in shRNA-1 group were (7.03±3.36)% and (9.40±2.51)%, respectively, which were decreased significantly compared with those of the neg-shRNA group(17.70±4.39)% and (22.63±1.86)%, and the control group (16.57±2.30)% and(21.67±1.93)% (P<0.05). Conclusions The shRNA targeting FOXP3 gene could efficiently inhibit the gene expression and hence it could significantly inhibit the apoptosis of glioma cells and promote cell proliferation. Key words: Glioma; Forkhead box P3; Apoptosis; Proliferation