Effect of pre-B-cell leukemia homeobox 3 gene small interfering RNA on apoptosis of glioma cells in vitro
Yongming Li, Yajun Zhao, Tao Qin
Abstract
Yongming Li, Yajun Zhao, Tao Qin
Abstract
Objective To investigate the effect of pre-B-cell leukemia homeobox 3 (PBX3) gene small interfering RNA (siRNA) on the viability and apoptosis of glioma cells. Methods According to Lipofectamine™ 2000, the specific siRNA of PBX3 (PBX3-siRNA group) was transfected into human glioma U87 cells, and the negative control siRNA (negative group) was transfected, and the blank group was set up, cell was transfected for 48 h, the expression of p38, p-p38, p53, cysteinyl aspartate specific proteinase-3 (Caspase-3) and cleaved Caspase-3 protein were detected by Western blotting. The cell apoptosis rate was detected by flow cytometry. Methyl thiazol tetrazolium (MTT) was used to detect the cell viability after PBX3-siRNA was transfected cells for 24, 48 and 72 h. Results The expression of PBX3 in U87 cells transfected with PBX3-siRNA was significantly lower than that in blank group (Protein relative expression were 0.096±0.010 and 0.467±0.051, P=0.000). Compared with the blank group, the cell viability in PBX3-siRNA group (24, 48, 72 h were 0.336±0.034, 0.549±0.057, 0.671±0.073, P24 h=0.001, P48 h=0.004, P72 h=0.004) decreased significantly PBX3-siRNA was transfected for 24-72 h, the apoptosis rate increased significantly, the expression of p-p38 protein(0.031±0.006) decreased significantly, and the expression of p53(0.388±0.041) and Cleaved-Caspase-3 protein (0.205±0.018) increased significantly (Pp-p38=0.000, Pp53=0.000, PCleaved-Caspase-3=0.000). There was no significant difference in the expression of p38 and Caspase-3 between the three groups (Pp38=0.987, PCaspase-3=0.871). Conclusion PBX3 gene siRNA can inhibit glioma cell viability and induce apoptosis, which may be related to downregulation of p38MAPK signal. Key words: Glioma; Pre-B-cell leukemia homeobox 3 gene; Apoptosis; p38 mitogen-activated protein kinases signal
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Objective To investigate the effect of pre-B-cell leukemia homeobox 3 (PBX3) gene small interfering RNA (siRNA) on the viability and apoptosis of glioma cells. Methods According to Lipofectamine™ 2000, the specific siRNA of PBX3 (PBX3-siRNA group) was transfected into human glioma U87 cells, and the negative control siRNA (negative group) was transfected, and the blank group was set up, cell was transfected for 48 h, the expression of p38, p-p38, p53, cysteinyl aspartate specific proteinase-3 (Caspase-3) and cleaved Caspase-3 protein were detected by Western blotting. The cell apoptosis rate was detected by flow cytometry. Methyl thiazol tetrazolium (MTT) was used to detect the cell viability after PBX3-siRNA was transfected cells for 24, 48 and 72 h. Results The expression of PBX3 in U87 cells transfected with PBX3-siRNA was significantly lower than that in blank group (Protein relative expression were 0.096±0.010 and 0.467±0.051, P=0.000). Compared with the blank group, the cell viability in PBX3-siRNA group (24, 48, 72 h were 0.336±0.034, 0.549±0.057, 0.671±0.073, P24 h=0.001, P48 h=0.004, P72 h=0.004) decreased significantly PBX3-siRNA was transfected for 24-72 h, the apoptosis rate increased significantly, the expression of p-p38 protein(0.031±0.006) decreased significantly, and the expression of p53(0.388±0.041) and Cleaved-Caspase-3 protein (0.205±0.018) increased significantly (Pp-p38=0.000, Pp53=0.000, PCleaved-Caspase-3=0.000). There was no significant difference in the expression of p38 and Caspase-3 between the three groups (Pp38=0.987, PCaspase-3=0.871). Conclusion PBX3 gene siRNA can inhibit glioma cell viability and induce apoptosis, which may be related to downregulation of p38MAPK signal. Key words: Glioma; Pre-B-cell leukemia homeobox 3 gene; Apoptosis; p38 mitogen-activated protein kinases signal
Key concepts: Transfection, Molecular biology, Viability assay, Apoptosis, Small interfering RNA, Lipofectamine, Cell, Biology